ANTIBODY THERAPIES FOR HUMAN IMMUNODEFICIENCY VIRUS (HIV)
20220025021 · 2022-01-27
Inventors
- Dan H. Barouch (Newton, MA)
- Bruce A. Kerwin (Bainbridge Island, WA)
- Randal R. Ketchem (Snohomish, WA)
- Alison J. Gillespie (Seattle, WA)
- Christine C. Siska (Seattle, WA)
- Rutilio H. Clark (Bainbridge Island, WA, US)
- Julee A. Floyd (Seattle, WA)
- Jeremy M. SHAVER (Lake Forest Park, WA, US)
Cpc classification
A61K39/39575
HUMAN NECESSITIES
A61K45/06
HUMAN NECESSITIES
C07K16/1063
CHEMISTRY; METALLURGY
A61K39/39575
HUMAN NECESSITIES
C07K2317/94
CHEMISTRY; METALLURGY
C07K2317/76
CHEMISTRY; METALLURGY
A61K2300/00
HUMAN NECESSITIES
A61K2300/00
HUMAN NECESSITIES
C07K2317/92
CHEMISTRY; METALLURGY
International classification
Abstract
Featured are PGDM1400 variant antibodies or fragments thereof, which can be administered, e.g., as antibody therapies for treating human immunodeficiency virus (HIV) infection. In particular, featured are methods of treating or curing subjects infected with HIV and/or preventing HIV infections in subjects at risk of HIV transmission using the PGDM1400 variant antibodies or fragments thereof.
Claims
1. An antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable domain comprising a sequence with at least 85% sequence identity to SEQ ID NO: 136; and (b) a light chain variable domain comprising a sequence with at least 85% sequence identity to SEQ ID NO: 135, wherein the antibody or antigen-binding fragment thereof comprises: (i) at least one of the following mutations in the heavy chain variable domain sequence: HV:P25S, HV:N27Y, HV:L29F, HV:Q46E, HV:D71T, HV:W72R, HV:Q82E, HV:T87R, and HV:D113E; and/or (ii) at least one of the following mutations in the light chain variable domain sequence: KV:F2I, KV:H9L, KV:S12P, KV:S18P, KV:R47Q, KV:D73G, KV:K74T, KV:T85A, and KV:T90V.
2. The antibody or antigen-binding fragment thereof of claim 1, wherein: (a) the heavy chain variable domain sequence has at least 85% sequence identity to SEQ ID NO: 136; and (b) the light chain variable domain sequence has at least 85% sequence identity to SEQ ID NO: 135 and at least one of the following mutations: KV:F2I, KV:H9L, KV:S12P, KV:S18P, KV:R47Q, KV:D73G, KV:K74T, KV:T85A, and KV:T90V.
3. The antibody or antigen-binding fragment thereof of claim 1, wherein: (a) the heavy chain variable domain sequence has at least 85% sequence identity to SEQ ID NO: 136 and at least one of the following mutations: HV:P25S, HV:N27Y, HV:L29F, HV:Q46E, HV:D71T, HV:W72R, HV:Q82E, HV:T87R, and HV:D113E; and (b) the light chain variable domain has at least 85% sequence identity to SEQ ID NO: 135.
4. The antibody or antigen-binding fragment thereof of claim 1, wherein: (a) the heavy chain variable domain sequence has at least 85% sequence identity to SEQ ID NO: 136 and at least one of the following mutations: HV:P25S, HV:N27Y, HV:L29F, HV:Q46E, HV:D71T, HV:W72R, HV:Q82E, HV:T87R, and HV:D113E; and (b) the light chain variable domain has at least 85% sequence identity to SEQ ID NO: 135 and at least one of the following mutations: KV:F2I, KV:H9L, KV:S12P, KV:S18P, KV:R47Q, KV:D73G, KV:K74T, KV:T85A, and KV:T90V.
5. The antibody or antigen-binding fragment thereof of any one of claims 1-4, further comprising an Fc domain comprising the amino acid sequence of SEQ ID NO: 137.
6. The antibody or antigen-binding fragment thereof of any one of claims 1-4, further comprising an Fc domain comprising the amino acid sequence of SEQ ID NO: 138.
7. The antibody or antigen-binding fragment thereof of any one of claims 1-6, wherein the antibody is a V2-specific antibody.
8. The antibody or antigen-binding fragment thereof of any one of claims 1-7, wherein the antibody or antigen-binding fragment thereof comprises: (a) (i) a heavy chain (HC) complementarity determining region (CDR) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a light chain (LC)-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 144 or amino acids 20-238 of SEQ ID NO: 18; (b) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 145 or amino acids 20-238 of SEQ ID NO: 20; (c) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 146 or amino acids 20-238 of SEQ ID NO: 22; (d) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 147 or amino acids 20-238 of SEQ ID NO: 24; (e) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 148 or amino acids 20-238 of SEQ ID NO: 26; (f) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 149 or amino acids 20-238 of SEQ ID NO: 28; (g) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 150 or amino acids 20-238 of SEQ ID NO: 30; (h) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 151 or amino acids 20-238 of SEQ ID NO: 32; (i) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 152 or amino acids 20-238 of SEQ ID NO: 34; (j) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 153 or amino acids 20-490 of SEQ ID NO: 36, and a light chain variable domain comprising the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2; (k) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 154 or amino acids 20-490 of SEQ ID NO: 38, and a light chain variable domain comprising the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2; (l) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 155 or amino acids 20-490 of SEQ ID NO: 40, and a light chain variable domain comprising the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2; (m) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 156 or amino acids 20-490 of SEQ ID NO: 42, and a light chain variable domain comprising the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2; (n) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 157 or amino acids 20-490 of SEQ ID NO: 44, and a light chain variable domain comprising the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2; (o) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 158 or amino acids 20-490 of SEQ ID NO: 46, and a light chain variable domain comprising the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2; (p) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 159 or amino acids 20-490 of SEQ ID NO: 48, and a light chain variable domain comprising the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2; (q) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 160 or amino acids 20-490 of SEQ ID NO: 50, and a light chain variable domain comprising the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2; (r) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 54, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 161 or amino acids 20-490 of SEQ ID NO: 52, and a light chain variable domain comprising the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2; (s) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 163 or amino acids 20-490 of SEQ ID NO: 58, and a light chain variable domain comprising the sequence of SEQ ID NO: 162 or amino acids 20-238 of SEQ ID NO: 56; (t) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 164 or amino acids 20-238 of SEQ ID NO: 60; (u) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 165 or amino acids 20-490 of SEQ ID NO: 62, and a light chain variable domain comprising the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2; (v) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 166 or amino acids 20-490 of SEQ ID NO: 64, and a light chain variable domain comprising the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2; (w) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 167 or amino acids 20-490 of SEQ ID NO: 66, and a light chain variable domain comprising the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2; (x) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 169 or amino acids 20-490 of SEQ ID NO: 70, and a light chain variable domain comprising the sequence of SEQ ID NO: 168 or amino acids 20-238 of SEQ ID NO: 68; (y) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 171 or amino acids 20-490 of SEQ ID NO: 74, and a light chain variable domain comprising the sequence of SEQ ID NO: 170 or amino acids 20-238 of SEQ ID NO: 72; (z) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 173 or amino acids 20-490 of SEQ ID NO: 78, and a light chain variable domain comprising the sequence of SEQ ID NO: 172 or amino acids 20-238 of SEQ ID NO: 76; (aa) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 175 or amino acids 20-490 of SEQ ID NO: 82, and a light chain variable domain comprising the sequence of SEQ ID NO: 174 or amino acids 20-238 of SEQ ID NO: 80; (bb) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2; (cc) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 176 or amino acids 20-238 of SEQ ID NO: 84; (dd) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 177 or amino acids 20-238 of SEQ ID NO: 86; (ee) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 178 or amino acids 20-238 of SEQ ID NO: 88; (ff) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 179 or amino acids 20-238 of SEQ ID NO: 90; (gg) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 180 or amino acids 20-238 of SEQ ID NO: 92; (hh) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 181 or amino acids 20-238 of SEQ ID NO: 94; (ii) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 182 or amino acids 20-238 of SEQ ID NO: 96; (jj) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 183 or amino acids 20-238 of SEQ ID NO: 98; (kk) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 184 or amino acids 20-238 of SEQ ID NO: 100; (ll) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 185 or amino acids 20-238 of SEQ ID NO: 102; (mm) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 186 or amino acids 20-238 of SEQ ID NO: 104; (nn) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 187 or amino acids 20-238 of SEQ ID NO: 106; (oo) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 188 or amino acids 20-238 of SEQ ID NO: 108; (pp) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 189 or amino acids 20-238 of SEQ ID NO: 110; (qq) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 190 or amino acids 20-238 of SEQ ID NO: 112; (rr) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 191 or amino acids 20-238 of SEQ ID NO: 114; (ss) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 192 or amino acids 20-238 of SEQ ID NO: 116; (tt) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 193 or amino acids 20-238 of SEQ ID NO: 118; (uu) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 194 or amino acids 20-238 of SEQ ID NO: 120; (vv) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 195 or amino acids 20-238 of SEQ ID NO: 122; (ww) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 196 or amino acids 20-238 of SEQ ID NO: 124; (xx) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 197 or amino acids 20-238 of SEQ ID NO: 126; (yy) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 198 or amino acids 20-238 of SEQ ID NO: 128; (zz) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 199 or amino acids 20-238 of SEQ ID NO: 130; (aaa) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 200 or amino acids 20-238 of SEQ ID NO: 132; or (bbb) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 201 or amino acids 20-238 of SEQ ID NO: 134.
9. The antibody or antigen-binding fragment thereof of claim 8, wherein the antibody or antigen-binding fragment thereof is selected from the group consisting of (a), (b), (d), (f), (h), (cc), (dd), (ee), (ff), (gg), (hh), (ii), (jj), (kk), (ll), (mm), (nn), (oo), (pp), (qq), (rr), (ss), (tt), (uu), (vv), (ww), (xx), (yy), (zz), (aaa), and (bbb).
10. The antibody or antigen-binding fragment thereof of claim 9, wherein the antibody or antigen-binding fragment thereof is selected from the group consisting of (cc), (dd), (ee), (ff), (gg), (hh), (ii), (jj), (kk), (ll), (mm), (nn), (oo), (pp), (qq), (rr), (ss), (tt), (uu), (vv), (ww), (xx), (yy), (zz), (aaa), and (bbb).
11. The antibody or antigen-binding fragment thereof of claim 10, wherein the antibody or antigen-binding fragment thereof is selected from the group consisting of (cc), (dd), (ee), (ff), (mm), (nn), (oo), (pp), (qq), (rr), (ww), (xx), (yy), (zz), and (bbb).
12. The antibody or antigen-binding fragment thereof of claim 11, wherein the antibody or antigen-binding fragment thereof is (cc).
13. The antibody or antigen-binding fragment thereof of any one of claims 1-12, wherein the antibody or antigen-binding fragment thereof exhibits one or more of the following properties: (i) neutralization of one or more of the following pseudoviruses of human immunodeficiency virus (HIV): SC422661.8, RHPA4259.7, Du172.17, BB1012-11.TC21, CNE52, 0260.v5.c36, 263-8, SC05.8C11.2344, X1193_c1, Cell 76_A3, AC10.0.29, and 6952.v1.c20; (ii) increased solubility, wherein optionally the antibody or antigen-binding fragment thereof is soluble in a PEG 10,000 concentration of 6-10%; (iii) increased stability at low pH, wherein optionally the low pH is less than pH 5.0; (iv) increased thermal stability; wherein optionally the antibody or antigen-binding fragment thereof is stable at a temperature in the range of 20−95° C.; and/or (v) increased chemical stability, wherein optionally the antibody or antigen-binding fragment thereof is resistant to chemical denaturation by guanidine hydrochloride (GuHCl), such as amount of GuHCl greater than 2 M, as compared to an antibody or antigen-binding fragment thereof lacking the at least one mutation in the heavy chain variable domain and/or the light chain variable domain.
14. The antibody or antigen-binding fragment thereof of claim 13, wherein the PEG 10,000 concentration is about 9.4%.
15. The antibody or antigen-binding fragment thereof of claim 13, wherein the temperature is about 68° C. or about 69.2° C.
16. The antibody or antigen-binding fragment thereof of claim 13, wherein the low pH is about pH 3.3.
17. The antibody or antigen-binding fragment thereof of claim 13, wherein the amount of GuHCl is about 6.0 M.
18. The antibody or antigen-binding fragment thereof of any one of claims 1-17, wherein the antibody or antigen-binding fragment thereof has increased storage stability.
19. The antibody or antigen-binding fragment thereof of claim 18, wherein the antibody or antigen-binding fragment thereof does not aggregate during storage over a period of time, wherein preferentially the time is over about 2 days.
20. The antibody or antigen-binding fragment thereof of any one of claims 1-19, wherein the antibody or antigen-binding fragment thereof has increased manufacturability.
21. The antibody or antigen-binding fragment thereof of claim 20, wherein the antibody or antigen-binding fragment thereof does not aggregate during manufacture.
22. The antibody or antigen-binding fragment thereof of any one of claims 18-21, wherein the antibody or antigen-binding fragment thereof exhibits high monomer content and/or low oligomer content.
23. The antibody or antigen-binding fragment thereof of claim 22, wherein the antibody or antigen-binding fragment thereof exhibits more than about 60% monomer content.
24. The antibody or antigen-binding fragment thereof of claim 22, wherein the antibody or antigen-binding fragment thereof exhibits less than about 10% oligomer content.
25. The antibody or antigen-binding fragment thereof of any one of claims 1-24, wherein the antibody or antigen-binding fragment thereof has a half-life in a fluid of at least 1 hour in vitro or in vivo.
26. The antibody or antigen-binding fragment thereof of claim 25, wherein the fluid is blood.
27. The antibody or antigen-binding fragment thereof of any one of claims 1-26, wherein the antibody or antigen-binding fragment thereof is selected from the group consisting of a monoclonal antibody or antigen-binding fragment thereof, a polyclonal antibody or antigen-binding fragment thereof, a human antibody or antigen-binding fragment thereof, a humanized antibody or antigen-binding fragment thereof, a primatized antibody or antigen-binding fragment thereof, a bispecific antibody or antigen-binding fragment thereof, a multi-specific antibody or antigen-binding fragment thereof, a dual-variable immunoglobulin domain, a monovalent antibody or antigen-binding fragment thereof, a chimeric antibody or antigen-binding fragment thereof, a single-chain Fv molecule (scFv), a diabody, a triabody, a nanobody, an antibody-like protein scaffold, a domain antibody, a Fv fragment, a Fab fragment, a F(ab′).sub.2 molecule, and a tandem scFv (taFv).
28. A polynucleotide encoding the antibody or antigen-binding fragment thereof of any one of claims 1-27.
29. A vector comprising the polynucleotide of claim 28.
30. The vector of claim 29, wherein the vector is an expression vector.
31. The vector of claim 30, wherein the expression vector is a prokaryotic or eukaryotic expression vector.
32. The vector of claim 29, wherein the vector is a viral vector.
33. The vector of claim 32, wherein the viral vector is selected from the group consisting of an adenovirus (Ad), a retrovirus, a poxvirus, an adeno-associated virus, a baculovirus, a herpes simplex virus, and a vaccinia virus.
34. The vector of claim 33, wherein the adenovirus is a serotype 2, 5, 11, 12, 24, 26, 34, 35, 40, 48, 49, 50, 52, or Pan9 adenovirus, or a human, chimpanzee, or rhesus adenovirus.
35. The vector of claim 33, wherein the retrovirus is a γ-retrovirus or a lentivirus.
36. The vector of claim 33, wherein the vaccinia virus is a modified vaccinia Ankara (MVA).
37. An isolated host cell comprising the polynucleotide of claim 28 or the vector of any one of claims 29-36.
38. The isolated host cell of claim 37, wherein the host cell is a prokaryotic cell or a eukaryotic cell.
39. The host cell of claim 38, wherein the eukaryotic cell is a mammalian cell.
40. The host cell of claim 39, wherein the mammalian cell is a Chinese Hamster Ovary (CHO) cell or a Human Embryonic Kidney 293 (HEK293) cell.
41. A composition comprising the antibody or antigen-binding fragment thereof of any one of claims 1-27, the polynucleotide of claim 28, the vector of any one of claims 29-36, or the host cell of any one of claims 37-40.
42. The composition of claim 41, further comprising a pharmaceutically acceptable carrier, excipient, or diluent.
43. The composition of claim 41 or 42, further comprising an immunomodulator.
44. The composition of claim 43, wherein the immunomodulator is one or more of AS-101, Bropirimine, Acemannan, CL246,738, EL10, FP-21399, Gamma Interferon, Granulocyte Macrophage Colony Stimulating Factor, HIV Core Particle Immunostimulant, IL-2, Immune Globulin Intravenous, IMREG-1, IMREG-2, Imuthiol Diethyl Dithio Carbamate, Alpha-2 Interferon, Methionine-Enkephalin, MTP-PE Muramyl-Tripeptide, Granulocyte Colony Stimulating Factor, Remune, CD4 such as recombinant soluble CD4, rCD4-IgG hybrids, SK&F106528 Soluble T4, Thymopentin, Tumor Necrosis Factor, and Infliximab.
45. The composition of any one of claims 41-44, further comprising at least one reservoir activator.
46. The composition of claim 45, wherein the reservoir activator is a PKC agonist, a cytokine or chemokine, a Toll-like receptor (TLR) agonist, an immune checkpoint inhibitor, a histone deacetylase (HDAC) inhibitor, or a small molecule reservoir activator.
47. The composition of claim 46, wherein: (a) the PKC agonist comprises one or more of a phorbol ester; a macrocyclic lactone, such as bryostatin-1; and/or a diterpene, such as an ingenol compound; (b) the cytokine or chemokine comprises one or more of interleukin (IL)-7, IL-15, or interferon-alpha (IFN-α); (c) the TLR agonist comprises one or more of a TLR 1/2 agonist, such as Pam3CSK4; a TLR3 agonist, such as Poly-ICLC; a TLR5 agonist, such as flagellin; a TLR7 agonist, such as GS-9620; and/or a TLR9 agonist, such as MGN1703 and CpG7909; (d) the immune checkpoint inhibitor comprises one or more of an anti-PD-1 monoclonal antibody; an anti-PD-1 ligand (PD-L1) monoclonal antibody; and/or an anti-CTLA-4 monoclonal antibody; (e) the HDAC inhibitor comprises one or more of romidepsin; vorinostat; belinostat; LAQ824; panobinostat; entinostat; C1994; and/or mocetinostat; (f) the small molecule reservoir activator comprises one or more of disulfiram; a benzotriazole derivative, such as 3-Hydroxy-1,2,3-benzotriazin-4((3H)-one (HO-DHBt); a SMAC mimetic; or a BRG-Brahma Associated Factor (BAF) inhibitor, such as caffeic acid phenethyl ester or pyrimethamine.
48. The composition of any one of claims 41-47, further comprising an antiretroviral agent (ARV).
49. The composition of claim 48, wherein the ARV comprises one or more of lamivudine and zidovudine, emtricitabine (FTC), zidovudine (ZDV), azidothymidine (AZT), lamivudine (3TC), zalcitabine, dideoxycytidine (ddC), tenofovir disoproxil fumarate (TDF), didanosine (ddl), stavudine (d4T), abacavir sulfate (ABC), etravirine, delavirdine (DLV), efavirenz (EFV), nevirapine (NVP), amprenavir (APV), tipranavir (TPV), indinavir (IDV), saquinavir, saquinavir mesylate (SQV), lopinavir (LPV), ritonavir (RTV), fosamprenavir calcium (FOS-APV), ritonavir, RTV, darunavir, atazanavir sulfate (ATV), nelfinavir mesylate (NFV), enfuvirtide, T-20, maraviroc, raltegravir, ibalizumab, IL-2, IL-12, or alpha-epibromide.
50. The composition of any one of claims 41-49, further comprising one, two, three, or more different HIV-specific broadly neutralizing antibodies (bnAb).
51. The composition of claim 50, wherein the bnAb is a CD4 binding site (CD4bs)-specific antibody or a V2 glycan-dependent antibody.
52. The composition of claim 51, wherein: (a) the CD4bs-specific antibody is 3BNC117 or VRC07-523, preferably wherein the CD4bs-specific antibody is 3BNC117; and/or (b) the V2 glycan dependent antibody is CAP256-VRC26.
53. The composition of any one of claims 41-52, wherein the composition comprises the antibody or antigen-binding fragment thereof in an amount of about 0.01-5000 mg.
54. The composition of any one of claims 41-53, wherein the composition is formulated for subcutaneous, intramuscular, intradermal, transdermal, intranasal, or oral administration, or administration as an infusion, wherein optionally the infusion is a continuous infusion or a bolus infusion.
55. The composition of any one of claims 41-54, wherein the composition is formulated in a volume of about 1000 ml or less.
56. The composition of claim 55, wherein the composition is formulated in a volume between about 0.1-1 ml.
57. A method of treating or blocking an HIV infection in a subject comprising administering to the subject the antibody or antigen-binding fragment thereof of any one of claims 1-27 or the composition of any one of claims 41-56.
58. The method of claim 57, wherein the antibody or antigen-binding fragment thereof or the composition is administered to the subject in a dosage form.
59. The method of claim 58, wherein about 0.01-5000 mg of the antibody or antigen-binding fragment thereof is administered to the subject.
60. The method of claim 58, wherein about 0.01-100 mg/kg of the antibody or antigen-binding fragment thereof is administered to the subject.
61. The method of any one of claims 57-60, wherein the antibody or antigen-binding fragment thereof is administered to the subject two or more times.
62. The method of claim 61, wherein the antibody or antigen-binding fragment thereof is administered to the subject one or more times daily, weekly, every two weeks, every three weeks, or monthly.
63. The method of any one of claims 57-62 wherein a single dose of the antibody or antigen-binding fragment thereof is administered to the subject.
64. The method of any one of claims 57-62, wherein more than one dose of the antibody or antigen-binding fragment thereof is administered to the subject.
65. The method of claim 64, wherein a second dose of the antibody or antigen-binding fragment thereof is administered to the subject two weeks, or more after administration of the first dose.
66. The method of any one of claims 57-65, wherein the subject is administered the antibody or antigen-binding fragment thereof for at least one week, or more.
67. The method of any one of claims 57-66, wherein administration of the antibody or antigen-binding fragment thereof reduces proviral DNA in a tissue of the subject relative to an untreated control.
68. The method of claim 67, wherein administration of the antibody or antigen-binding fragment thereof reduces the proviral DNA in the tissue to below about 1,000 DNA copies/10.sup.6 cells.
69. The method of any one of claims 57-68, wherein the administration of the antibody or antigen-binding fragment thereof reduces the proviral DNA in the tissue to an undetectable level.
70. The method of claim 69, wherein HIV therapy is concluded when the administration of the antibody or antigen-binding fragment thereof reduces the proviral DNA in the tissue to an undetectable level.
71. The method of any one of claims 67-70, wherein the tissue is lymph node tissue, gastrointestinal tissue, and/or peripheral blood.
72. The method of any one of claims 57-71, wherein the subject has a plasma viral load of less than 3,500 RNA copies/ml following administration of the antibody or antigen-binding fragment thereof.
73. The method of any one of claims 57-72, wherein the subject has an undetectable plasma viral load following administration of the antibody or antigen-binding fragment thereof.
74. The method of claim 73, wherein the subject has an undetectable plasma viral load for at least 2 months following administration of the antibody or antigen-binding fragment thereof.
75. The method of any one of claims 57-74, wherein the administration of the antibody or antigen-binding fragment thereof increases HIV-specific cell-mediated immune response and/or humoral immune response in the subject relative to an untreated control.
76. The method of any one of claims 57-75, wherein the administration of the antibody or antigen-binding fragment thereof decreases viral replication in the subject relative to an untreated control.
77. The method of any one of claims 57-76, wherein the antibody or antigen-binding fragment thereof is administered intravenously, intramuscularly, intradermally, percutaneously, intraarterially, intraperitoneally, intralesionally, intracranially, intraarticularly, intraprostatically, intrapleurally, intratracheally, intranasally, intravitreally, intravaginally, intrarectally, topically, intratumorally, peritoneally, subcutaneously, subconjunctivally, intravesicularlly, mucosally, intrapericardially, intraumbilically, intraocularly, orally, topically, locally, by inhalation, by injection, by infusion, by continuous infusion, by localized perfusion bathing target cells directly, by catheter, by lavage, by gavage, in cremes, or in lipid compositions.
78. The method of any one of claims 57-77, wherein the antibody or antigen-binding fragment thereof is administered in combination with one or more immunomodulators, reservoir activators, ARVs, and/or HIV-specific bnAb.
79. The method of claim 78, wherein the immunomodulator is one or more of AS-101, Bropirimine, Acemannan, CL246,738, EL10, FP-21399, Gamma Interferon, Granulocyte Macrophage Colony Stimulating Factor, HIV Core Particle Immunostimulant, IL-2, Immune Globulin Intravenous, IMREG-1, IMREG-2, Imuthiol Diethyl Dithio Carbamate, Alpha-2 Interferon, Methionine-Enkephalin, MTP-PE Muramyl-Tripeptide, Granulocyte Colony Stimulating Factor, Remune, CD4 (e.g., recombinant soluble CD4), rCD4-IgG hybrids, SK&F106528 Soluble T4, Thymopentin, Tumor Necrosis Factor, or Infliximab.
80. The method of claim 78, wherein the reservoir activator is a PKC agonist, a cytokine or chemokine, a Toll-like receptor (TLR) agonist, an immune checkpoint inhibitor, a histone deacetylase (HDAC) inhibitor, or a small molecule reservoir activator.
81. The method of claim 80, wherein: (a) the PKC agonist comprises one or more of a phorbol ester; a macrocyclic lactone, such as bryostatin-1; and/or a diterpene, such as an ingenol compound; (b) the cytokine or chemokine comprises one or more of interleukin (IL)-7, IL-15, or interferon-alpha (IFN-α); (c) the TLR agonist comprises one or more of a TLR 1/2 agonist, such as Pam3CSK4; a TLR3 agonist, such as Poly-ICLC; a TLR5 agonist, such as flagellin; a TLR7 agonist, such as GS-9620; and/or a TLR9 agonist, such as MGN1703 and CpG7909; (d) the immune checkpoint inhibitor comprises one or more of an anti-PD-1 monoclonal antibody; an anti-PD-1 ligand (PD-L1) monoclonal antibody; and/or an anti-CTLA-4 monoclonal antibody; (e) the HDAC inhibitor comprises one or more of romidepsin; vorinostat; belinostat; LAQ824; panobinostat; entinostat; C1994; and/or mocetinostat; (f) the small molecule reservoir activator comprises one or more of disulfiram; a benzotriazole derivative, such as 3-Hydroxy-1,2,3-benzotriazin-4((3H)-one (HO-DHBt); a SMAC mimetic; or a BRG-Brahma Associated Factor (BAF) inhibitor, such as caffeic acid phenethyl ester or pyrimethamine.
82. The method of claim 78, wherein the ARV comprises one or more of lamivudine and zidovudine, emtricitabine (FTC), zidovudine (ZDV), azidothymidine (AZT), lamivudine (3TC), zalcitabine, dideoxycytidine (ddC), tenofovir disoproxil fumarate (TDF), didanosine (ddl), stavudine (d4T), abacavir sulfate (ABC), etravirine, delavirdine (DLV), efavirenz (EFV), nevirapine (NVP), amprenavir (APV), tipranavir (TPV), indinavir (IDV), saquinavir, saquinavir mesylate (SQV), lopinavir (LPV), ritonavir (RTV), fosamprenavir calcium (FOS-APV), ritonavir, RTV, darunavir, atazanavir sulfate (ATV), nelfinavir mesylate (NFV), enfuvirtide, T-20, maraviroc, raltegravir, ibalizumab, IL-2, IL-12, or alpha-epibromide.
83. The method of claim 78, wherein the bnAb is a CD4 binding site (CD4bs)-specific antibody or an N332 glycan dependent antibody.
84. The method of claim 83, wherein: (a) the CD4bs-specific antibody is 3BNC117 or VRC07-523, preferably wherein said CD4bs-specific antibody is 3BNC117; and/or (b) the N332 glycan dependent antibody is PGT121.
85. The method of any one of claims 78-84, wherein the immunomodulator, the reservoir activator, the ARV, and/or the HIV-specific bnAb is/are administered prior to, concurrently, and/or after the administration of the antibody or antigen-binding fragment thereof.
86. The method of claim 85, wherein the immunomodulator, the reservoir activator, the ARV, and/or the HIV-specific bnAb is/are administered: (a) 1 hour, or more prior to the administration of the antibody or antigen-binding fragment thereof; (b) concurrent to the administration of the antibody or antigen-binding fragment thereof; and/or (c) 1 hour, or more after the administration of the antibody or antigen-binding fragment thereof.
87. The method of any one of claims 57-86, wherein: (a) the subject is infected with HIV; or (b) the subject is at risk of HIV transmission.
88. The method of claim 87, wherein the subject at risk of HIV transmission is: (a) a fetus of an HIV-infected pregnant female; (b) a newborn having an HIV-infected mother; (c) a subject having a needle stick injury; (d) a subject being sexually exposed to one or more HIV-infected individuals.
89. The method of any one of claims 57-88, wherein the subject is a human.
90. The method of any one of claims 57-89, wherein the HIV infection is an HIV type 1 (HIV-1) and/or an HIV type 2 (HIV-2) infection.
91. The method of claim 90, wherein the HIV infection is an HIV-1 infection.
92. A kit comprising the antibody or antigen-binding fragment thereof of any one of claims 1-27, the polynucleotide of claim 28, the vector of any one of claims 29-36, the host cell of any one of claims 37-40, or the composition of any one of claims 41-56 in a therapeutically effective amount for preventing or treating HIV infection in a subject according to the method of any one of claims 57-91.
93. The kit of claim 92 further comprising instructions, wherein the instructions are for the purpose of directing a clinician in methods for administering to the subject the antibody or antigen-binding fragment thereof, the polynucleotide, the vector, the host cell or the composition contained therein.
94. The antibody or antigen-binding fragment thereof of claim 1, wherein the antibody is a V2-specific antibody.
95. The antibody or antigen-binding fragment thereof of claim 1, wherein the antibody or antigen-binding fragment thereof comprises: (a) (i) a heavy chain (HC) complementarity determining region (CDR) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a light chain (LC)-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 144 or amino acids 20-238 of SEQ ID NO: 18; (b) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 145 or amino acids 20-238 of SEQ ID NO: 20; (c) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 146 or amino acids 20-238 of SEQ ID NO: 22; (d) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 147 or amino acids 20-238 of SEQ ID NO: 24; (e) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 148 or amino acids 20-238 of SEQ ID NO: 26; (f) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 149 or amino acids 20-238 of SEQ ID NO: 28; (g) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 150 or amino acids 20-238 of SEQ ID NO: 30; (h) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 151 or amino acids 20-238 of SEQ ID NO: 32; (i) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 152 or amino acids 20-238 of SEQ ID NO: 34; (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 153 or amino acids 20-490 of SEQ ID NO: 36, and a light chain variable domain comprising the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2; (k) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 154 or amino acids 20-490 of SEQ ID NO: 38, and a light chain variable domain comprising the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2; (l) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 155 or amino acids 20-490 of SEQ ID NO: 40, and a light chain variable domain comprising the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2; (m) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 156 or amino acids 20-490 of SEQ ID NO: 42, and a light chain variable domain comprising the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2; (n) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 157 or amino acids 20-490 of SEQ ID NO: 44, and a light chain variable domain comprising the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2; (o) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 158 or amino acids 20-490 of SEQ ID NO: 46, and a light chain variable domain comprising the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2; (p) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 159 or amino acids 20-490 of SEQ ID NO: 48, and a light chain variable domain comprising the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2; (q) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 160 or amino acids 20-490 of SEQ ID NO: 50, and a light chain variable domain comprising the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2; (r) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 54, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 161 or amino acids 20-490 of SEQ ID NO: 52, and a light chain variable domain comprising the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2; (s) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 163 or amino acids 20-490 of SEQ ID NO: 58, and a light chain variable domain comprising the sequence of SEQ ID NO: 162 or amino acids 20-238 of SEQ ID NO: 56; (t) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 164 or amino acids 20-238 of SEQ ID NO: 60; (u) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 165 or amino acids 20-490 of SEQ ID NO: 62, and a light chain variable domain comprising the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2; (v) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 166 or amino acids 20-490 of SEQ ID NO: 64, and a light chain variable domain comprising the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2; (w) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 167 or amino acids 20-490 of SEQ ID NO: 66, and a light chain variable domain comprising the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2; (x) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 169 or amino acids 20-490 of SEQ ID NO: 70, and a light chain variable domain comprising the sequence of SEQ ID NO: 168 or amino acids 20-238 of SEQ ID NO: 68; (y) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 171 or amino acids 20-490 of SEQ ID NO: 74, and a light chain variable domain comprising the sequence of SEQ ID NO: 170 or amino acids 20-238 of SEQ ID NO: 72; (z) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 173 or amino acids 20-490 of SEQ ID NO: 78, and a light chain variable domain comprising the sequence of SEQ ID NO: 172 or amino acids 20-238 of SEQ ID NO: 76; (aa) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 175 or amino acids 20-490 of SEQ ID NO: 82, and a light chain variable domain comprising the sequence of SEQ ID NO: 174 or amino acids 20-238 of SEQ ID NO: 80; (bb) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2; (cc) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 176 or amino acids 20-238 of SEQ ID NO: 84; (dd) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 177 or amino acids 20-238 of SEQ ID NO: 86; (ee) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 178 or amino acids 20-238 of SEQ ID NO: 88; (ff) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 179 or amino acids 20-238 of SEQ ID NO: 90; (gg) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 180 or amino acids 20-238 of SEQ ID NO: 92; (hh) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 181 or amino acids 20-238 of SEQ ID NO: 94; (ii) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 182 or amino acids 20-238 of SEQ ID NO: 96; (jj) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 183 or amino acids 20-238 of SEQ ID NO: 98; (kk) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 184 or amino acids 20-238 of SEQ ID NO: 100; (ll) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 185 or amino acids 20-238 of SEQ ID NO: 102; (mm) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 186 or amino acids 20-238 of SEQ ID NO: 104; (nn) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 187 or amino acids 20-238 of SEQ ID NO: 106; (oo) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 188 or amino acids 20-238 of SEQ ID NO: 108; (pp) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 189 or amino acids 20-238 of SEQ ID NO: 110; (qq) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 190 or amino acids 20-238 of SEQ ID NO: 112; (rr) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 191 or amino acids 20-238 of SEQ ID NO: 114; (ss) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 192 or amino acids 20-238 of SEQ ID NO: 116; (tt) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 193 or amino acids 20-238 of SEQ ID NO: 118; (uu) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 194 or amino acids 20-238 of SEQ ID NO: 120; (vv) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 195 or amino acids 20-238 of SEQ ID NO: 122; (ww) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 196 or amino acids 20-238 of SEQ ID NO: 124; (xx) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 197 or amino acids 20-238 of SEQ ID NO: 126; (yy) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 198 or amino acids 20-238 of SEQ ID NO: 128; (zz) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 199 or amino acids 20-238 of SEQ ID NO: 130; (aaa) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 200 or amino acids 20-238 of SEQ ID NO: 132; or (bbb) (i) a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16, a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; and/or (ii) a heavy chain variable domain comprising the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain comprising the sequence of SEQ ID NO: 201 or amino acids 20-238 of SEQ ID NO: 134.
96. The antibody or antigen-binding fragment thereof of claim 95, wherein the antibody or antigen-binding fragment thereof is selected from the group consisting of (a), (b), (d), (f), (h), (cc), (dd), (ee), (ff), (gg), (hh), (ii), (jj), (kk), (ll), (mm), (nn), (oo), (pp), (qq), (rr), (ss), (tt), (uu), (vv), (ww), (xx), (yy), (zz), (aaa), and (bbb).
97. The antibody or antigen-binding fragment thereof of claim 96, wherein the antibody or antigen-binding fragment thereof is selected from the group consisting of (cc), (dd), (ee), (ff), (gg), (hh), (ii), (jj), (kk), (ll), (mm), (nn), (oo), (pp), (qq), (rr), (ss), (tt), (uu), (vv), (ww), (xx), (yy), (zz), (aaa), and (bbb).
98. The antibody or antigen-binding fragment thereof of claim 97, wherein the antibody or antigen-binding fragment thereof is selected from the group consisting of (cc), (dd), (ee), (ff), (mm), (nn), (oo), (pp), (qq), (rr), (ww), (xx), (yy), (zz), and (bbb).
99. The antibody or antigen-binding fragment thereof of claim 98, wherein the antibody or antigen-binding fragment thereof is (cc).
100. The antibody or antigen-binding fragment thereof of claim 1, wherein the antibody or antigen-binding fragment thereof exhibits one or more of the following properties: (i) neutralization of one or more of the following pseudoviruses of human immunodeficiency virus (HIV): SC422661.8, RHPA4259.7, Du172.17, BB1012-11.TC21, CNE52, 0260.v5.c36, 263-8, SC05.8C11.2344, X1193_c1, Cell 76_A3, AC10.0.29, and 6952.v1.c20; (ii) increased solubility, wherein optionally the antibody or antigen-binding fragment thereof is soluble in a PEG 10,000 concentration of 6-10%; (iii) increased stability at low pH, wherein optionally the low pH is less than pH 5.0; (iv) increased thermal stability; wherein optionally the antibody or antigen-binding fragment thereof is stable at a temperature in the range of 20-95° C.; and/or (v) increased chemical stability, wherein optionally the antibody or antigen-binding fragment thereof is resistant to chemical denaturation by guanidine hydrochloride (GuHCl), such as amount of GuHCl greater than 2 M, as compared to an antibody or antigen-binding fragment thereof lacking the at least one mutation in the heavy chain variable domain and/or the light chain variable domain.
101. The antibody or antigen-binding fragment thereof of claim 100, wherein the PEG 10,000 concentration is about 9.4%.
102. The antibody or antigen-binding fragment thereof of claim 100, wherein the temperature is about 68° C. or about 69.2° C.
103. The antibody or antigen-binding fragment thereof of claim 100, wherein the low pH is about pH 3.3.
104. The antibody or antigen-binding fragment thereof of claim 100, wherein the amount of GuHCl is about 6.0 M.
105. The antibody or antigen-binding fragment thereof of claim 1, wherein the antibody or antigen-binding fragment thereof has increased storage stability.
106. The antibody or antigen-binding fragment thereof of claim 105, wherein the antibody or antigen-binding fragment thereof does not aggregate during storage over a period of time, wherein preferentially the time is over about 2 days.
107. The antibody or antigen-binding fragment thereof of claim 1, wherein the antibody or antigen-binding fragment thereof has increased manufacturability.
108. The antibody or antigen-binding fragment thereof of claim 107, wherein the antibody or antigen-binding fragment thereof does not aggregate during manufacture.
109. The antibody or antigen-binding fragment thereof of claim 105, wherein the antibody or antigen-binding fragment thereof exhibits high monomer content and/or low oligomer content.
110. The antibody or antigen-binding fragment thereof of claim 109, wherein the antibody or antigen-binding fragment thereof exhibits more than about 60% monomer content.
111. The antibody or antigen-binding fragment thereof of claim 109, wherein the antibody or antigen-binding fragment thereof exhibits less than about 10% oligomer content.
112. The antibody or antigen-binding fragment thereof of claim 1, wherein the antibody or antigen-binding fragment thereof has a half-life in a fluid of at least 1 hour in vitro or in vivo.
113. The antibody or antigen-binding fragment thereof of claim 112, wherein the fluid is blood.
114. The antibody or antigen-binding fragment thereof of claim 1, wherein the antibody or antigen-binding fragment thereof is selected from the group consisting of a monoclonal antibody or antigen-binding fragment thereof, a polyclonal antibody or antigen-binding fragment thereof, a human antibody or antigen-binding fragment thereof, a humanized antibody or antigen-binding fragment thereof, a primatized antibody or antigen-binding fragment thereof, a bispecific antibody or antigen-binding fragment thereof, a multi-specific antibody or antigen-binding fragment thereof, a dual-variable immunoglobulin domain, a monovalent antibody or antigen-binding fragment thereof, a chimeric antibody or antigen-binding fragment thereof, a single-chain Fv molecule (scFv), a diabody, a triabody, a nanobody, an antibody-like protein scaffold, a domain antibody, a Fv fragment, a Fab fragment, a F(ab′).sub.2 molecule, and a tandem scFv (taFv).
115. A polynucleotide encoding the antibody or antigen-binding fragment thereof of claim 1.
116. A vector comprising the polynucleotide of claim 115.
117. The vector of claim 116, wherein the vector is an expression vector.
118. The vector of claim 117, wherein the expression vector is a prokaryotic or eukaryotic expression vector.
119. The vector of claim 116, wherein the vector is a viral vector.
120. The vector of claim 119, wherein the viral vector is selected from the group consisting of an adenovirus (Ad), a retrovirus, a poxvirus, an adeno-associated virus, a baculovirus, a herpes simplex virus, and a vaccinia virus.
121. The vector of claim 120, wherein the adenovirus is a serotype 2, 5, 11, 12, 24, 26, 34, 35, 40, 48, 49, 50, 52, or Pan9 adenovirus, or a human, chimpanzee, or rhesus adenovirus.
122. The vector of claim 120, wherein the retrovirus is a γ-retrovirus or a lentivirus.
123. The vector of claim 120, wherein the vaccinia virus is a modified vaccinia Ankara (MVA).
124. An isolated host cell comprising the polynucleotide of claim 115 or the vector of claim 116.
125. The isolated host cell of claim 124, wherein the host cell is a prokaryotic cell or a eukaryotic cell.
126. The host cell of claim 125, wherein the eukaryotic cell is a mammalian cell.
127. The host cell of claim 126, wherein the mammalian cell is a Chinese Hamster Ovary (CHO) cell or a Human Embryonic Kidney 293 (HEK293) cell.
128. A composition comprising the antibody or antigen-binding fragment thereof of claim 1, the polynucleotide of claim 115, the vector of claim 116, or the host cell of claim 124.
129. The composition of claim 128, further comprising a pharmaceutically acceptable carrier, excipient, or diluent.
130. The composition of claim 128 or 129, further comprising an immunomodulator.
131. The composition of claim 130, wherein the immunomodulator is one or more of AS-101, Bropirimine, Acemannan, CL246,738, EL10, FP-21399, Gamma Interferon, Granulocyte Macrophage Colony Stimulating Factor, HIV Core Particle Immunostimulant, IL-2, Immune Globulin Intravenous, IMREG-1, IMREG-2, Imuthiol Diethyl Dithio Carbamate, Alpha-2 Interferon, Methionine-Enkephalin, MTP-PE Muramyl-Tripeptide, Granulocyte Colony Stimulating Factor, Remune, CD4 such as recombinant soluble CD4, rCD4-IgG hybrids, SK&F106528 Soluble T4, Thymopentin, Tumor Necrosis Factor, and Infliximab.
132. The composition of claim 128, further comprising at least one reservoir activator.
133. The composition of claim 132, wherein the reservoir activator is a PKC agonist, a cytokine or chemokine, a Toll-like receptor (TLR) agonist, an immune checkpoint inhibitor, a histone deacetylase (HDAC) inhibitor, or a small molecule reservoir activator.
134. The composition of claim 133, wherein: (a) the PKC agonist comprises one or more of a phorbol ester; a macrocyclic lactone, such as bryostatin-1; and/or a diterpene, such as an ingenol compound; (b) the cytokine or chemokine comprises one or more of interleukin (IL)-7, IL-15, or interferon-alpha (IFN-α); (c) the TLR agonist comprises one or more of a TLR 1/2 agonist, such as Pam3CSK4; a TLR3 agonist, such as Poly-ICLC; a TLR5 agonist, such as flagellin; a TLR7 agonist, such as GS-9620; and/or a TLR9 agonist, such as MGN1703 and CpG7909; (d) the immune checkpoint inhibitor comprises one or more of an anti-PD-1 monoclonal antibody; an anti-PD-1 ligand (PD-L1) monoclonal antibody; and/or an anti-CTLA-4 monoclonal antibody; (e) the HDAC inhibitor comprises one or more of romidepsin; vorinostat; belinostat; LAQ824; panobinostat; entinostat; 01994; and/or mocetinostat; (f) the small molecule reservoir activator comprises one or more of disulfiram; a benzotriazole derivative, such as 3-Hydroxy-1,2,3-benzotriazin-4((3H)-one (HO-DHBt); a SMAC mimetic; or a BRG-Brahma Associated Factor (BAF) inhibitor, such as caffeic acid phenethyl ester or pyrimethamine.
135. The composition of claim 128, further comprising an antiretroviral agent (ARV).
136. The composition of claim 135, wherein the ARV comprises one or more of lamivudine and zidovudine, emtricitabine (FTC), zidovudine (ZDV), azidothymidine (AZT), lamivudine (3TC), zalcitabine, dideoxycytidine (ddC), tenofovir disoproxil fumarate (TDF), didanosine (ddl), stavudine (d4T), abacavir sulfate (ABC), etravirine, delavirdine (DLV), efavirenz (EFV), nevirapine (NVP), amprenavir (APV), tipranavir (TPV), indinavir (IDV), saquinavir, saquinavir mesylate (SQV), lopinavir (LPV), ritonavir (RTV), fosamprenavir calcium (FOS-APV), ritonavir, RTV, darunavir, atazanavir sulfate (ATV), nelfinavir mesylate (NFV), enfuvirtide, T-20, maraviroc, raltegravir, ibalizumab, IL-2, IL-12, or alpha-epibromide.
137. The composition of claim 128, further comprising one, two, three, or more different HIV-specific broadly neutralizing antibodies (bnAb).
138. The composition of claim 137, wherein the bnAb is a CD4 binding site (CD4bs)-specific antibody or a V2 glycan-dependent antibody.
139. The composition of claim 138, wherein: (a) the CD4bs-specific antibody is 3BNC117 or VRC07-523, preferably wherein the CD4bs-specific antibody is 3BNC117; and/or (b) the V2 glycan dependent antibody is CAP256-VRC26.
140. The composition of claim 128, wherein the composition comprises the antibody or antigen-binding fragment thereof in an amount of about 0.01-5000 mg.
141. The composition of claim 128, wherein the composition is formulated for subcutaneous, intramuscular, intradermal, transdermal, intranasal, or oral administration, or administration as an infusion, wherein optionally the infusion is a continuous infusion or a bolus infusion.
142. The composition of claim 128, wherein the composition is formulated in a volume of about 1000 ml or less.
143. The composition of claim 142, wherein the composition is formulated in a volume between about 0.1-1 ml.
144. A method of treating or blocking an HIV infection in a subject comprising administering to the subject the antibody or antigen-binding fragment thereof of claim 1 or the composition of claim 128.
145. The method of claim 144, wherein the antibody or antigen-binding fragment thereof or the composition is administered to the subject in a dosage form.
146. The method of claim 145, wherein about 0.01-5000 mg of the antibody or antigen-binding fragment thereof is administered to the subject.
147. The method of claim 145, wherein about 0.01-100 mg/kg of the antibody or antigen-binding fragment thereof is administered to the subject.
148. The method of claim 144, wherein the antibody or antigen-binding fragment thereof is administered to the subject two or more times.
149. The method of claim 148, wherein the antibody or antigen-binding fragment thereof is administered to the subject one or more times daily, weekly, every two weeks, every three weeks, or monthly.
150. The method of claim 144 wherein a single dose of the antibody or antigen-binding fragment thereof is administered to the subject.
151. The method of claim 144, wherein more than one dose of the antibody or antigen-binding fragment thereof is administered to the subject.
152. The method of claim 151, wherein a second dose of the antibody or antigen-binding fragment thereof is administered to the subject two weeks, or more after administration of the first dose.
153. The method of claim 144, wherein the subject is administered the antibody or antigen-binding fragment thereof for at least one week, or more.
154. The method of claim 144, wherein administration of the antibody or antigen-binding fragment thereof reduces proviral DNA in a tissue of the subject relative to an untreated control.
155. The method of claim 154, wherein administration of the antibody or antigen-binding fragment thereof reduces the proviral DNA in the tissue to below about 1,000 DNA copies/10.sup.6 cells.
156. The method of claim 144, wherein the administration of the antibody or antigen-binding fragment thereof reduces the proviral DNA in the tissue to an undetectable level.
157. The method of claim 156, wherein HIV therapy is concluded when the administration of the antibody or antigen-binding fragment thereof reduces the proviral DNA in the tissue to an undetectable level.
158. The method of claim 154, wherein the tissue is lymph node tissue, gastrointestinal tissue, and/or peripheral blood.
159. The method of claim 144, wherein the subject has a plasma viral load of less than 3,500 RNA copies/ml following administration of the antibody or antigen-binding fragment thereof.
160. The method of claim 144, wherein the subject has an undetectable plasma viral load following administration of the antibody or antigen-binding fragment thereof.
161. The method of claim 160, wherein the subject has an undetectable plasma viral load for at least 2 months following administration of the antibody or antigen-binding fragment thereof.
162. The method of claim 144, wherein the administration of the antibody or antigen-binding fragment thereof increases HIV-specific cell-mediated immune response and/or humoral immune response in the subject relative to an untreated control.
163. The method of claim 144, wherein the administration of the antibody or antigen-binding fragment thereof decreases viral replication in the subject relative to an untreated control.
164. The method of claim 144, wherein the antibody or antigen-binding fragment thereof is administered intravenously, intramuscularly, intradermally, percutaneously, intraarterially, intraperitoneally, intralesionally, intracranially, intraarticularly, intraprostatically, intrapleurally, intratracheally, intranasally, intravitreally, intravaginally, intrarectally, topically, intratumorally, peritoneally, subcutaneously, subconjunctivally, intravesicularlly, mucosally, intrapericardially, intraumbilically, intraocularly, orally, topically, locally, by inhalation, by injection, by infusion, by continuous infusion, by localized perfusion bathing target cells directly, by catheter, by lavage, by gavage, in cremes, or in lipid compositions.
165. The method of claim 144, wherein the antibody or antigen-binding fragment thereof is administered in combination with one or more immunomodulators, reservoir activators, ARVs, and/or HIV-specific bnAb.
166. The method of claim 165, wherein the immunomodulator is one or more of AS-101, Bropirimine, Acemannan, CL246,738, EL10, FP-21399, Gamma Interferon, Granulocyte Macrophage Colony Stimulating Factor, HIV Core Particle Immunostimulant, IL-2, Immune Globulin Intravenous, IMREG-1, IMREG-2, Imuthiol Diethyl Dithio Carbamate, Alpha-2 Interferon, Methionine-Enkephalin, MTP-PE Muramyl-Tripeptide, Granulocyte Colony Stimulating Factor, Remune, CD4 (e.g., recombinant soluble CD4), rCD4-IgG hybrids, SK&F106528 Soluble T4, Thymopentin, Tumor Necrosis Factor, or Infliximab.
167. The method of claim 165, wherein the reservoir activator is a PKC agonist, a cytokine or chemokine, a Toll-like receptor (TLR) agonist, an immune checkpoint inhibitor, a histone deacetylase (HDAC) inhibitor, or a small molecule reservoir activator.
168. The method of claim 167, wherein: (a) the PKC agonist comprises one or more of a phorbol ester; a macrocyclic lactone, such as bryostatin-1; and/or a diterpene, such as an ingenol compound; (b) the cytokine or chemokine comprises one or more of interleukin (IL)-7, IL-15, or interferon-alpha (IFN-α); (c) the TLR agonist comprises one or more of a TLR 1/2 agonist, such as Pam3CSK4; a TLR3 agonist, such as Poly-ICLC; a TLR5 agonist, such as flagellin; a TLR7 agonist, such as GS-9620; and/or a TLR9 agonist, such as MGN1703 and CpG7909; (d) the immune checkpoint inhibitor comprises one or more of an anti-PD-1 monoclonal antibody; an anti-PD-1 ligand (PD-L1) monoclonal antibody; and/or an anti-CTLA-4 monoclonal antibody; (e) the HDAC inhibitor comprises one or more of romidepsin; vorinostat; belinostat; LAQ824; panobinostat; entinostat; C1994; and/or mocetinostat; (f) the small molecule reservoir activator comprises one or more of disulfiram; a benzotriazole derivative, such as 3-Hydroxy-1,2,3-benzotriazin-4((3H)-one (HO-DHBt); a SMAC mimetic; or a BRG-Brahma Associated Factor (BAF) inhibitor, such as caffeic acid phenethyl ester or pyrimethamine.
169. The method of claim 165, wherein the ARV comprises one or more of lamivudine and zidovudine, emtricitabine (FTC), zidovudine (ZDV), azidothymidine (AZT), lamivudine (3TC), zalcitabine, dideoxycytidine (ddC), tenofovir disoproxil fumarate (TDF), didanosine (ddl), stavudine (d4T), abacavir sulfate (ABC), etravirine, delavirdine (DLV), efavirenz (EFV), nevirapine (NVP), amprenavir (APV), tipranavir (TPV), indinavir (IDV), saquinavir, saquinavir mesylate (SQV), lopinavir (LPV), ritonavir (RTV), fosamprenavir calcium (FOS-APV), ritonavir, RTV, darunavir, atazanavir sulfate (ATV), nelfinavir mesylate (NFV), enfuvirtide, T-20, maraviroc, raltegravir, ibalizumab, IL-2, IL-12, or alpha-epibromide.
170. The method of claim 165, wherein the bnAb is a CD4 binding site (CD4bs)-specific antibody or an N332 glycan dependent antibody.
171. The method of claim 170, wherein: (a) the CD4bs-specific antibody is 3BNC117 or VRC07-523, preferably wherein said CD4bs-specific antibody is 3BNC117; and/or (b) the N332 glycan dependent antibody is PGT121.
172. The method of claim 165, wherein the immunomodulator, the reservoir activator, the ARV, and/or the HIV-specific bnAb is/are administered prior to, concurrently, and/or after the administration of the antibody or antigen-binding fragment thereof.
173. The method of claim 172, wherein the immunomodulator, the reservoir activator, the ARV, and/or the HIV-specific bnAb is/are administered: (a) 1 hour, or more prior to the administration of the antibody or antigen-binding fragment thereof; (b) concurrent to the administration of the antibody or antigen-binding fragment thereof; and/or (c) 1 hour, or more after the administration of the antibody or antigen-binding fragment thereof.
174. The method of claim 144, wherein: (a) the subject is infected with HIV; or (b) the subject is at risk of HIV transmission.
175. The method of claim 174, wherein the subject at risk of HIV transmission is: (a) a fetus of an HIV-infected pregnant female; (b) a newborn having an HIV-infected mother; (c) a subject having a needle stick injury; (d) a subject being sexually exposed to one or more HIV-infected individuals.
176. The method of claim 144, wherein the subject is a human.
177. The method of claim 144, wherein the HIV infection is an HIV type 1 (HIV-1) and/or an HIV type 2 (HIV-2) infection.
178. The method of claim 177, wherein the HIV infection is an HIV-1 infection.
179. A kit comprising the antibody or antigen-binding fragment thereof of claim 1, the polynucleotide of claim 115, the vector of claim 116, the host cell of claim 124, or the composition of claim 128 in a therapeutically effective amount for preventing or treating HIV infection in a subject according to the method of claim 144.
180. The kit of claim 179 further comprising instructions, wherein the instructions are for the purpose of directing a clinician in methods for administering to the subject the antibody or antigen-binding fragment thereof, the polynucleotide, the vector, the host cell or the composition contained therein.
Description
BRIEF DESCRIPTION OF DRAWINGS
[0178]
[0179]
[0180]
[0181]
[0182]
DETAILED DESCRIPTION OF THE INVENTION
[0183] We have identified and mutated potentially destabilizing residues in the variable domain (Fv) of the PGDM1400 antibody. These residues of the antibody, by themselves or in combination, may lead to instability at low pH, increased susceptibility to chemical degradation, or aggregation during production or long term storage. Based on our discovery, we generated a series of antibody variants with mutations of one or more of the destabilizing residues. The antibody variants produced by such combinatorial residue replacement techniques retained potency (e.g., viral inactivation or neutralization potency) while exhibiting desired biophysical characteristics, in particular, increased stability at low pH, reduced susceptibility to chemical degradation, and reduced aggregation. Featured herein are PGDM1400 variant antibodies and antigen-binding fragments thereof that retain the ability of the native PGDM1400 antibody to inactivate or neutralize viruses (e.g., HIV-1), while showing significant improvement in production efficiency (e.g., increased production titer), manufacturability, and storage stability relative to the native PGDM1400 antibody.
I. Antibodies and Antigen-Binding Fragments Thereof
[0184] Featured are PGDM1400 variant antibodies and antigen-binding fragments thereof that exhibit improved properties. The PGDM1400 variant antibodies or fragment thereof contain: (a) a heavy chain variable domain having a sequence with at least 85% (e.g., at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO: 136; and (b) a light chain variable domain having a sequence with at least 85% (e.g., at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO: 135; and wherein the antibody or antigen-binding fragment thereof has: (i) at least one of the following mutations in the heavy chain variable domain sequence: HV:P25S, HV:N27Y, HV:L29F, HV:Q46E, HV:D71T, HV:W72R, HV:Q82E, HV:T87R, and HV:D113E; and/or (ii) at least one of the following mutations in the light chain variable domain sequence: KV:F2I, KV:H9L, KV:S12P, KV:S18P, KV:R47Q, KV:D73G, KV:K74T, KV:T85A, and KV:T90V.
[0185] For example, the PGDM1400 variant antibody or fragment thereof may contain (i) a heavy chain variable domain having a sequence with at least 85% sequence identity to SEQ ID NO: 136; and (ii) a light chain variable domain having a sequence with at least 85% sequence identity to SEQ ID NO: 135, and at least one (e.g., at least one, at least two, at least three, at least four, at least five, at least six, or more) of the following mutations in the light chain variable domain: KV:F2I, KV:H9L, KV:S12P, KV:S18P, KV:R47Q, KV:D73G, KV:K74T, KV:T85A, and KV:T90V. Alternatively, the PGDM1400 variant antibody or fragment thereof may have (i) a heavy chain variable domain having a sequence with at least 85% sequence identity to SEQ ID NO: 136, and at least one (e.g., at least one, at least two, at least three, at least four, at least five, at least six, or more) of the following mutations in the heavy chain variable domain: HV:P25S, HV:N27Y, HV:L29F, HV:Q46E, HV:D71T, HV:W72R, HV:Q82E, HV:T87R, and HV:D113E; and (ii) a light chain variable domain having a sequence with at least 85% sequence identity to SEQ ID NO: 135. In some embodiments, the PGDM1400 variant antibody or fragment thereof may have (i) a heavy chain variable domain having a sequence with at least 85% sequence identity to SEQ ID NO: 136, and at least one (e.g., at least one, at least two, at least three, at least four, at least five, at least six, or more) of the following mutations in the heavy chain variable domain: HV:P25S, HV:N27Y, HV:L29F, HV:Q46E, HV:D71T, HV:W72R, HV:Q82E, HV:T87R, and HV:D113E; and (ii) a light chain variable domain having a sequence with at least 85% sequence identity to SEQ ID NO: 135, and at least one (e.g., at least one, at least two, at least three, at least four, at least five, at least six, or more) of the following mutations in the light chain variable domain: KV:F2I, KV:H9L, KV:S12P, KV:S18P, KV:R47Q, KV:D73G, KV:K74T, KV:T85A, and KV:T90V. In other embodiments, the PGDM1400 variant antibody or fragment thereof may have (i) a heavy chain variable domain having a sequence with at least 85% sequence identity to SEQ ID NO: 136; (ii) a light chain variable domain having a sequence with at least 85% sequence identity to SEQ ID NO: 135; (iii) at least one (e.g., at least one, at least two, at least three, at least four, at least five, at least six, or more) of the following mutations in the heavy chain variable domain: HV:P25S, HV:N27Y, HV:L29F, HV:Q46E, HV:D71T, HV:W72R, HV:Q82E, HV:T87R, and HV:D113E; and (iv) at least one (e.g., at least one, at least two, at least three, at least four, at least five, at least six, or more) of the following mutations in the light chain variable domain: KV:F2I, KV:H9L, KV:S12P, KV:S18P, KV:R47Q, KV:D73G, KV:K74T, KV:T85A, and KV:T90V. Alternatively, the PGDM1400 variant antibody or fragment thereof may contain (i) a heavy chain variable domain having a sequence with at least 85% sequence identity to SEQ ID NO: 136; and (ii) a light chain variable domain having a sequence with at least 85% sequence identity to SEQ ID NO: 135.
[0186] In some embodiments, the PGDM1400 variant antibody or fragment thereof may have (i) a heavy chain variable domain having a sequence with at least 85% sequence identity to SEQ ID NO: 136; (ii) a light chain variable domain having a sequence with at least 85% sequence identity to SEQ ID NO: 135; and (iii) a KV:F2I mutation in the light chain variable domain. Such a PGDM1400 variant antibody or fragment thereof may further comprise: at least one (e.g., at least one, at least two, at least three, at least four, at least five, at least six, or more) of the following mutations in the heavy chain variable domain: HV:P25S, HV:N27Y, HV:L29F, HV:Q46E, HV:D71T, HV:W72R, HV:Q82E, HV:T87R, and HV:D113E; and/or at least one (e.g., at least one, at least two, at least three, at least four, at least five, at least six, or more) of the following mutations in the light chain variable domain: KV:H9L, KV:S12P, KV:S18P, KV:R47Q, KV:D73G, KV:K74T, KV:T85A, and KV:T90V.
[0187] The Fc domain of any of the PGDM1400 variant antibodies or fragments thereof described herein may include the sequence of SEQ ID NO: 137, or a sequence with at least 85% (e.g., at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO: 137. Alternatively, the Fc domain of any of the PGDM1400 variant antibodies or fragments thereof described herein may include the sequence of SEQ ID NO: 138, or a sequence with at least 85% (e.g., at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO: 138. Preferentially, the Fc domain of the PGDM1400 variant antibody or fragment thereof includes the sequence of SEQ ID NO: 138, or a sequence with at least 85% (e.g., at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO: 138. Alternatively, the Fc domain of the PGDM1400 variant antibody or fragment thereof may include a sequence with at least 85% (e.g., at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO: 137, and a M87L and/or a N93S mutation. The Fc domain of any of the PGDM1400 variant antibodies or fragments thereof described herein may further include the sequence of SEQ ID NO: 139, or a sequence with at least 85% (e.g., at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO: 139. Together, the Fc domain of any of the PGDM1400 variant antibodies or fragments thereof described herein may have: (i) the sequence of SEQ ID NO: 140, or a sequence with at least 85% (e.g., at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO: 140; or (ii) the sequence of SEQ ID NO: 141, or a sequence with at least 85% (e.g., at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO: 141.
[0188] The featured PGDM1400 variant antibody or fragment thereof may further include an Ig domain with the sequence of SEQ ID NO: 142, or a sequence with at least 85% (e.g., at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO: 142. Additionally, the antibody or antigen-binding fragment thereof described herein may further include a Hinge region with the sequence of SEQ ID NO: 143, or a sequence with at least 85% (e.g., at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO: 143.
[0189] In specific embodiments:
[0190] (a) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 144 or amino acids 20-238 of SEQ ID NO: 18. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and a KV:F2I mutation in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 17, respectively;
[0191] (b) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 145 or amino acids 20-238 of SEQ ID NO: 20. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and a KV:H9L mutation in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 19, respectively;
[0192] (c) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 146 or amino acids 20-238 of SEQ ID NO: 22. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and a KV:S12P mutation in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 21, respectively;
[0193] (d) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs):a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 147 or amino acids 20-238 of SEQ ID NO: 24. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and a KV:S18P mutation in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 23, respectively;
[0194] (e) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 148 or amino acids 20-238 of SEQ ID NO: 26. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and a KV:R47Q mutation in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 25, respectively;
[0195] (f) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 149 or amino acids 20-238 of SEQ ID NO: 28. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and a KV:D73G mutation in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 27, respectively;
[0196] (g) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 150 or amino acids 20-238 of SEQ ID NO: 30. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and a KV:K74T mutation in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 29, respectively;
[0197] (h) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 151 or amino acids 20-238 of SEQ ID NO: 32. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and a KV:T85A mutation in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 31, respectively;
[0198] (i) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 152 or amino acids 20-238 of SEQ ID NO: 34. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and a KV:T90V mutation in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 33, respectively;
[0199] (j) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 153 or amino acids 20-490 of SEQ ID NO: 36, and a light chain variable domain having the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2. The antibody or antigen-binding fragment thereof has a HV:P25S mutation in the heavy chain variable domain, and M87L and N93S mutations in the heavy chain Fc region. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 35, and 1, respectively;
[0200] (k) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 154 or amino acids 20-490 of SEQ ID NO: 38, and a light chain variable domain having the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2. The antibody or antigen-binding fragment thereof has a HV:N27Y mutation in the heavy chain variable domain, and M87L and N93S mutations in the heavy chain Fc region. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 37, and 1, respectively;
[0201] (l) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 155 or amino acids 20-490 of SEQ ID NO: 40, and a light chain variable domain having the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2. The antibody or antigen-binding fragment thereof has a HV:L29F mutation in the heavy chain variable domain, and M87L and N93S mutations in the heavy chain Fc region. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 39, and 1, respectively;
[0202] (m) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 156 or amino acids 20-490 of SEQ ID NO: 42, and a light chain variable domain having the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2. The antibody or antigen-binding fragment thereof has a HV:Q46E mutation in the heavy chain variable domain, and M87L and N93S mutations in the heavy chain Fc region. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 41, and 1, respectively;
[0203] (n) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 157 or amino acids 20-490 of SEQ ID NO: 44, and a light chain variable domain having the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2. The antibody or antigen-binding fragment thereof has a HV:D71T mutation in the heavy chain variable domain, and M87L and N93S mutations in the heavy chain Fc region. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 43, and 1, respectively;
[0204] (o) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 158 or amino acids 20-490 of SEQ ID NO: 46, and a light chain variable domain having the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2. The antibody or antigen-binding fragment thereof has a HV:W72R mutation in the heavy chain variable domain, and M87L and N93S mutations in the heavy chain Fc region. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 45, and 1, respectively;
[0205] (p) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 159 or amino acids 20-490 of SEQ ID NO: 48, and a light chain variable domain having the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2. The antibody or antigen-binding fragment thereof has a HV:Q82E mutation in the heavy chain variable domain, and M87L and N93S mutations in the heavy chain Fc region. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 47, and 1, respectively;
[0206] (q) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 160 or amino acids 20-490 of SEQ ID NO: 50, and a light chain variable domain having the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2. The antibody or antigen-binding fragment thereof has a HV:T87R mutation in the heavy chain variable domain, and M87L and N93S mutations in the heavy chain Fc region. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 49, and 1, respectively;
[0207] (r) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 54, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 54; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 161 or amino acids 20-490 of SEQ ID NO: 52, and a light chain variable domain having the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2. The antibody or antigen-binding fragment thereof has a HV:D113E mutation in the heavy chain variable domain, and M87L and N93S mutations in the heavy chain Fc region. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 53, 3, 5, 7, 51, and 1, respectively;
[0208] (s) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 163 or amino acids 20-490 of SEQ ID NO: 58, and a light chain variable domain having the sequence of SEQ ID NO: 162 or amino acids 20-238 of SEQ ID NO: 56. The antibody or antigen-binding fragment thereof has a HV:T87R mutation in the heavy chain variable domain, M87L and N93S mutations in the heavy chain Fc region, and KV:H9L, KV:S12P, KV:S18P, KV:R47Q, KV:T85A and KV:T90V mutations in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 57, and 55, respectively;
[0209] (t) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 164 or amino acids 20-238 of SEQ ID NO: 60. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and KV:F2I, KV:D73G and KV:K74T mutations in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 59, respectively;
[0210] (u) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 165 or amino acids 20-490 of SEQ ID NO: 62, and a light chain variable domain having the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2. The antibody or antigen-binding fragment thereof has HV:P25S, HV:N27Y and HV:L29F mutations in the heavy chain variable domain, and M87L and N93S mutations in the heavy chain Fc region. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 61, and 1, respectively;
[0211] (v) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 166 or amino acids 20-490 of SEQ ID NO: 64, and a light chain variable domain having the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2. The antibody or antigen-binding fragment thereof has HV:D71T and HV:W72R mutations in the heavy chain variable domain, and M87L and N93S mutations in the heavy chain Fc region. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 63, and 1, respectively;
[0212] (w) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 167 or amino acids 20-490 of SEQ ID NO: 66, and a light chain variable domain having the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2. The antibody or antigen-binding fragment thereof has HV:P25S, HV:N27Y, HV:L29F, HV:D71T and HV:W72R mutations in the heavy chain variable domain, and M87L and N93S mutations in the heavy chain Fc region. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 65, and 1, respectively;
[0213] (x) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 169 or amino acids 20-490 of SEQ ID NO: 70, and a light chain variable domain having the sequence of SEQ ID NO: 168 or amino acids 20-238 of SEQ ID NO: 68. The antibody or antigen-binding fragment thereof has HV:N27Y and HV:D71T mutations in the heavy chain variable domain, M87L and N93S mutations in the heavy chain Fc region, and a KV:H9L mutation in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 69, and 67, respectively;
[0214] (y) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 171 or amino acids 20-490 of SEQ ID NO: 74, and a light chain variable domain having the sequence of SEQ ID NO: 170 or amino acids 20-238 of SEQ ID NO: 72. The antibody or antigen-binding fragment thereof has HV:P25S, HV:N27Y and HV:L29F mutations in the heavy chain variable domain, M87L and N93S mutations in the heavy chain Fc region, and a KV:H9L mutation in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 73, and 71, respectively;
[0215] (z) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 173 or amino acids 20-490 of SEQ ID NO: 78, and a light chain variable domain having the sequence of SEQ ID NO: 172 or amino acids 20-238 of SEQ ID NO: 76. The antibody or antigen-binding fragment thereof has HV:P25S and HV:N27Y mutations in the heavy chain variable domain, M87L and N93S mutations in the heavy chain Fc region, and KV:H9L and KV:K74T mutations in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 77, and 75, respectively;
[0216] (aa) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; tor (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 175 or amino acids 20-490 of SEQ ID NO: 82, and a light chain variable domain having the sequence of SEQ ID NO: 174 or amino acids 20-238 of SEQ ID NO: 80. The antibody or antigen-binding fragment thereof has HV:Q46E, HV:W72R and HV:T87R mutations in the heavy chain variable domain, M87L and N93S mutations in the heavy chain Fc region, and a KV:F2I mutation in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 81, and 79, respectively;
[0217] (bb) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 135 or amino acids 20-238 of SEQ ID NO: 2. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 1, respectively;
[0218] (cc) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 176 or amino acids 20-238 of SEQ ID NO: 84. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and KV:F2I and KV:H9L mutations in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 83, respectively;
[0219] (dd) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 177 or amino acids 20-238 of SEQ ID NO: 86. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and KV:F2I and KV:S18P mutations in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 85, respectively;
[0220] (ee) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 178 or amino acids 20-238 of SEQ ID NO: 88. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and KV:F2I and KV:D73G mutations in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 87, respectively;
[0221] (ff) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 179 or amino acids 20-238 of SEQ ID NO: 90. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and KV:F2I and KV:T85A mutations in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 89, respectively;
[0222] (gg) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 180 or amino acids 20-238 of SEQ ID NO: 92. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and KV:H9L and KV:S18P mutations in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 91, respectively;
[0223] (hh) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 181 or amino acids 20-238 of SEQ ID NO: 94. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and KV:H9L and KV:D73G mutations in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 93, respectively;
[0224] (ii) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 182 or amino acids 20-238 of SEQ ID NO: 96. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and KV:H9L and KV:T85A mutations in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 95, respectively;
[0225] (jj) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 183 or amino acids 20-238 of SEQ ID NO: 98. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and KV:S18P and KV:D73G mutations in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 97, respectively;
[0226] (kk) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 184 or amino acids 20-238 of SEQ ID NO: 100. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and KV:S18P and KV:T85A mutations in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 99, respectively;
[0227] (ll) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 185 or amino acids 20-238 of SEQ ID NO: 102. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and KV:D73G and KV:T85A mutations in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 101, respectively;
[0228] (mm) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 186 or amino acids 20-238 of SEQ ID NO: 104. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and KV:F2I, KV:H9L and KV:S18P mutations in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 103, respectively;
[0229] (nn) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 187 or amino acids 20-238 of SEQ ID NO: 106. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and KV:F2I, KV:H9L and KV:D73G mutations in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 105, respectively;
[0230] (oo) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 188 or amino acids 20-238 of SEQ ID NO: 108. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and KV:F2I, KV:H9L and KV:T85A mutations in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 107, respectively;
[0231] (pp) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 189 or amino acids 20-238 of SEQ ID NO: 110. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and KV:F2I, KV:S18P and KV:D73G mutations in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 109, respectively;
[0232] (qq) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 190 or amino acids 20-238 of SEQ ID NO: 112. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and KV:F2I, KV:S18P and KV:T85A mutations in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 111, respectively;
[0233] (rr) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 191 or amino acids 20-238 of SEQ ID NO: 114. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and KV:F2I, KV:D73G and KV:T85A mutations in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 113, respectively;
[0234] (ss) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 192 or amino acids 20-238 of SEQ ID NO: 116. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and KV:H9L, KV:S18P and KV:D73G mutations in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 115, respectively;
[0235] (tt) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 193 or amino acids 20-238 of SEQ ID NO: 118. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and KV:H9L, KV:S18P and KV:T85A mutations in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 117, respectively;
[0236] (uu) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 194 or amino acids 20-238 of SEQ ID NO: 120. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and KV:H9L, KV:D73G and KV:T85A mutations in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 119, respectively;
[0237] (vv) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 195 or amino acids 20-238 of SEQ ID NO: 122. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and KV:S18P, KV:D73G and KV:T85A mutations in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 121, respectively;
[0238] (ww) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 196 or amino acids 20-238 of SEQ ID NO: 124. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and KV:F2I, KV:H9L, KV:S18P and KV:D73G mutations in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 123, respectively;
[0239] (xx) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 197 or amino acids 20-238 of SEQ ID NO: 126. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and KV:F2I, KV:H9L, KV:S18P and KV:T85A mutations in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 125, respectively;
[0240] (yy) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 198 or amino acids 20-238 of SEQ ID NO: 128. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and KV:F2I, KV:H9L, KV:D73G and KV:T85A mutations in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 127, respectively;
[0241] (zz) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 199 or amino acids 20-238 of SEQ ID NO: 130. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and KV:F2I, KV:S18P, KV:D73G and KV:T85A mutations in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 129, respectively;
[0242] (aaa) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 200 or amino acids 20-238 of SEQ ID NO: 132. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and KV:H9L, KV:S18P, KV:D73G and KV:T85A mutations in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 131, respectively; or (bbb) a PGDM1400 variant antibody or antigen-binding fragment thereof featured herein includes: (i) the following six complementarity determining regions (CDRs): a heavy chain (HC)-CDR1 with the amino acid sequence of SEQ ID NO: 12, or 3 or fewer (e.g., 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 12; a HC-CDR2 with the amino acid sequence of SEQ ID NO: 14, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 14; a HC-CDR3 with the amino acid sequence of SEQ ID NO: 16, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 16; a light chain (LC)-CDR1 with the amino acid sequence of SEQ ID NO: 4, or 10 or fewer (e.g., 9, 8, 7, 6, 5, 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 4; a LC-CDR2 with the amino acid sequence of SEQ ID NO: 6, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 6; and a LC-CDR3 with the amino acid sequence of SEQ ID NO: 8, or 5 or fewer (e.g., 4, 3, 2 or 1) amino acid modification(s) (e.g., insertion, deletion, or substitution) relative to the amino acid sequence of SEQ ID NO: 8; or (ii) a heavy chain variable domain having the sequence of SEQ ID NO: 136 or amino acids 20-490 of SEQ ID NO: 10, and a light chain variable domain having the sequence of SEQ ID NO: 201 or amino acids 20-238 of SEQ ID NO: 134. The antibody or antigen-binding fragment thereof has M87L and N93S mutations in the heavy chain Fc region, and KV:F2I, KV:H9L, KV:S18P, KV:D73G and KV:T85A mutations in the light chain variable domain. In a particular antibody or antigen-binding fragment thereof, the HC-CDR1, the HC-CDR2, the HC-CDR3, the LC-CDR1, the LC-CDR2, the LC-CDR3, the heavy chain variable domain, and the light chain variable domain of the antibody or antigen-binding fragment thereof are encoded by the nucleotide sequences of SEQ ID NOs: 11, 13, 15, 3, 5, 7, 9, and 133, respectively.
[0243] For manufacturing an antibody or antigen-binding fragment thereof of (a)-(bbb) above (e.g., using an expression system), the heavy and light chain amino acid sequences noted above may include a signal peptide. The signal peptide corresponds to residues 1-19 of the sequences noted above. During maturation, the signal peptide is cleaved. Hence, the mature form of the antibody or antigen-binding fragment thereof lacks the first 1-19 amino acids of the sequence of the respective heavy and light chain domain. The residue numbering corresponds to the amino acid position of the mature linear sequence for the heavy and light chain variable domains of the antibodies described herein, which excludes the signal peptide sequence (amino acids 1-19). For example, position 2 of the mature linear sequence of the light chain variable domain of MS-66 (i.e., SEQ ID NO: 144) begins at amino acid position 21 of SEQ ID NO: 18. Position 21 of SEQ ID NO: 18 corresponds to the KV:F2I substitution.
[0244] Residues 1-57 of the nucleotide sequence of heavy and light chain variable domains of the PGDM1400 variant antibody or antigen-binding fragment thereof featured herein (e.g., residue 1-57 of SEQ ID NOs: 1, 9, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, and 133) encode signal peptides, which, as noted in the foregoing section, are cleaved during maturation, and henceforth, are not a part of the mature linear sequence of the heavy and light chain variable domains of the PGDM1400 variant antibody or antigen-binding fragment thereof featured herein.
[0245] In specific embodiments, the PGDM1400 variant antibody or antigen-binding fragment thereof featured herein may be selected from the group consisting of the aforementioned: (a), (b), (d), (f), (h), (cc), (dd), (ee), (ff), (gg), (hh), (ii), (jj), (kk), (ll), (mm), (nn), (oo), (pp), (qq), (rr), (ss), (tt), (uu), (vv), (ww), (xx), (yy), (zz), (aaa), and (bbb). Specifically, the PGDM1400 variant antibody or antigen-binding fragment thereof featured herein may be selected from the group consisting of the aforementioned: (cc), (dd), (ee), (ff), (gg), (hh), (ii), (jj), (kk), (ll), (mm), (nn), (oo), (pp), (qq), (rr), (ss), (tt), (uu), (vv), (ww), (xx), (yy), (zz), (aaa), and (bbb). Preferentially, the PGDM1400 variant antibody or antigen-binding fragment thereof featured herein may be selected from the group consisting of the aforementioned: (cc), (dd), (ee), (ff), (mm), (nn), (oo), (pp), (qq), (rr), (ww), (xx), (yy), (zz), and (bbb). Preferably, the PGDM1400 variant antibody or antigen-binding fragment thereof featured herein is (cc) (e.g., MS-93).
[0246] In specific embodiments, the PGDM1400 variant antibody or antigen-binding fragment thereof featured herein may be selected from the group consisting of the following from Tables 1 and 2: MS-66, MS-67, MS-69, MS-71, MS-73, MS-93, MS-94, MS-95, MS-96, MS-97, MS-98, MS-99, MS-100, MS-101, MS-102, MS-103, MS-104, MS-105, MS-106, MS-107, MS-108, MS-109, MS-110, MS-111, MS-112, MS-113, MS-114, MS-115, MS-116, MS-117, and MS-118. In selective embodiments, the PGDM1400 variant antibody or antigen-binding fragment thereof featured herein may be selected from the group consisting of the following from Table 2: MS-93, MS-94, MS-95, MS-96, MS-97, MS-98, MS-99, MS-100, MS-101, MS-102, MS-103, MS-104, MS-105, MS-106, MS-107, MS-108, MS-109, MS-110, MS-111, MS-112, MS-113, MS-114, MS-115, MS-116, MS-117, and MS-118. Preferentially, the PGDM1400 variant antibody or antigen-binding fragment thereof featured herein may be selected from the group consisting of the following from Table 2: MS-93, MS-94, MS-95, MS-96, MS-103, MS-104, MS-105, MS-106, MS-107, MS-108, MS-113, MS-114, MS-115, MS-116, and MS-118. Preferably, the PGDM1400 variant antibody or antigen-binding fragment thereof featured herein is MS-93.
[0247] In some embodiments, the CDR sequences noted above for the PGDM1400 variant antibodies (a)-(bbb) may differ by one, two, three, four, five, six, seven, eight, nine, or ten amino acid residues from the recited sequences. In such embodiments, insertion (e.g., insertion of one, two, three, four, five, six, seven, eight, nine, or ten amino acid residues), deletion (e.g., deletion of one, two, three, four, five, six, seven, eight, nine, or ten amino acid residues), or substitution (e.g., substitution of one, two, three, four, five, six, seven, eight, nine, or ten amino acid residues) may account for the amino acid difference (e.g., difference of one, two, three, four, five, six, seven, eight, nine, or ten amino acid residues) of the CDR sequences from the recited CDR sequences noted herein. The amino acid substitution in the CDR(s), if present, may be a conservative amino acid substitution.
II. Design of the PGDM1400 Variant Antibodies
[0248] Antibody variants (e.g., PGDM1400 variant antibodies) or antigen-binding fragments thereof, described herein may be produced by an optimization process. The optimization process may be broken up into different stages with the first being identification of single residues in the framework region that may be responsible for destabilization of the parental PGDM1400 antibody. A series of variants can be produced by transient expression (e.g., transient expression in Human Embryonic Kidney 293 (HEK293) or Chinese Hamster Ovary (CHO) cells), each containing a single residue modification of amino acids, or in a few variants, combinations of amino acids based on proximity to each other (e.g., one or more of the Round-1 variants of Table 1). These variants may be characterized for retention of neutralization activity (e.g., neutralization activity against pseudoviruses of human immunodeficiency virus (HIV), such as SC422661.8, RHPA4259.7, Du172.17, BB1012-11.TC21, CNE52, 0260.v5.c36, 263-8, SC05.8C11.2344, X1193_c1, Ce1176_A3, AC10.0.29, and 6952.v1.c20) and for desired biophysical characteristics (e.g., low-pH stability, solubility, thermal stability, chemical unfolding, and reduced aggregation).
[0249] We identified several single residues at the light chain/heavy chain interface that significantly reduce low-pH instability (e.g., instability at pH 3.3) of the parental PGDM1400 antibody. Additionally, we identified amino acid residue combinations the substitution of which promoted an increase in desirable biophysical characteristics, while not impacting neutralization characteristics (e.g., neutralization or inactivation of viruses). Together, these residues were used to produce a library of variants (e.g., one or more of the Round-2 variants of Table 2) encompassing combinatorial residue replacements. The variants can be produced by transient expression (e.g., transient expression in HEK293 or CHO cells) and the purified combinatorial variants can be analyzed for retention of neutralization activity (e.g., neutralization activity against pseudoviruses of human immunodeficiency virus (HIV), such as SC422661.8, RHPA4259.7, Du172.17, BB1012-11.TC21, CNE52, 0260.v5.c36, 263-8, SC05.8C11.2344, X1193_c1, Ce1176_A3, AC10.0.29, and 6952.v1.c20) and for desired biophysical characteristics (e.g., low-pH stability, solubility, thermal stability, chemical unfolding, and reduced aggregation). From this combinatorial library a subset of variants may be used to construct a library. Together, the combinatorial libraries of variants allow for identification of antibody variants or fragments thereof with desired biophysical characteristics, such as with significantly increased low-pH stability (e.g., stability at about pH 3.3), increased thermal stability (e.g., tested during thermal ramping between about 20-95° C.), increased solubility (e.g., in a final PEG 10,000 concentration of about 9.4%), reduced aggregation (e.g., reduced levels of aggregation following low-pH (e.g., about pH 3.3) incubation) as evaluated by monomer and/or oligomer content (e.g., monomer content more than about 60% (e.g., more than about 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, or 97%), and/or oligomer content less than about 10% (e.g., less than about 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.4%, or 0.3%)), and increased intramolecular and thermodynamic stability, such as chemical stability, as determined by chemical unfolding (e.g., tested by guanidine hydrochloride (GuHCl) or urea concentrations, preferably by GuHCl concentrations).
TABLE-US-00001 TABLE 1 Round-1 variants. Light Chain (LC)-CDR 1-3 Light Chain Heavy Chain Amino acid (aa) SEQ ID NO: Variable Domain Variable Domain Nucleotide (nt) SEQ ID NO: Amino acid (aa) Amino acid (aa) IgG1 Light Chain IgG1 Heavy Chain Fc Domain Heavy Chain (HC)-CDR 1-3 SEQ ID NO: SEQ ID NO: Modification Modification Modification Molecule Amino acid (aa) SEQ ID NO: Nucleotide (nt) Nucleotide (nt) (Relative to (Relative to (Relative to Set Nucleotide (nt) SEQ ID NO: SEQ ID NO: SEQ ID NO: SEQ ID NO: 135) SEQ ID NO: 136) SEQ ID NO: 137) PGDM1400 LC-CDR 1-3 aa: 135 aa: 136 No modification No modification No modification aa: 4, 6, 8 nt: 1.sup.1 nt: 9 nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-119 LC-CDR 1-3 aa: 135 aa: 136 No modification No modification M87L; N93S aa: 4, 6, 8 nt: 1 nt: 9 nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-66 LC-CDR 1-3 aa: 144 aa: 136 KV: F2I No modification M87L; N93S aa: 4, 6, 8 nt: 17 nt: 9 nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-67 LC-CDR 1-3 aa: 145 aa: 136 KV: H9L No modification M87L; N93S aa: 4, 6, 8 nt: 19 nt: 9 nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-68 LC-CDR 1-3 aa: 146 aa: 136 KV: S12P No modification M87L; N93S aa: 4, 6, 8 nt: 21 nt: 9 nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-69 LC-CDR 1-3 aa: 147 aa: 136 KV: S18P No modification M87L; N93S aa: 4, 6, 8 nt: 23 nt: 9 nt: 3, 5, 7 HC-CDR 1-3 aa: −12, 14, 16 nt: 11, 13, 15 MS-70 LC-CDR 1-3 aa: 148 aa: 136 KV: R47Q No modification M87L; N93S aa: 4, 6, 8 nt: 25 nt: 9 nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-71 LC-CDR 1-3 aa: 149 aa: 136 KV: D73G No modification M87L; N93S aa: 4, 6, 8 nt: 27 nt: 9 nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-72 LC-CDR 1-3 aa: 150 aa: 136 KV: K74T No modification M87L; N93S aa: 4, 6, 8 nt: 29 nt: 9 nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-73 LC-CDR 1-3 aa: 151 aa: 136 KV: T85A No modification M87L; N93S aa: 4, 6, 8 nt: 31 nt: 9 nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-74 LC-CDR 1-3 aa: 152 aa: 136 KV: T90V No modification M87L; N93S aa: 4, 6, 8 nt: 33 nt: 9 nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-75 LC-CDR 1-3 aa: 135 aa: 153 No modification HV: P25S M87L; N93S aa: 4, 6, 8 nt: 1 nt: 35 nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-76 LC-CDR 1-3 aa: 135 aa: 154 No modification HV: N27Y M87L; N93S aa: 4, 6, 8 nt: 1 nt: 37 nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-77 LC-CDR 1-3 aa: 135 aa: 155 No modification HV: L29F M87L; N93S aa: 4, 6, 8 nt: 1 nt: 39 nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-78 LC-CDR 1-3 aa: 135 aa: 156 No modification HV: Q46E M87L; N93S aa: 4, 6, 8 nt: 1 nt: 41 nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-79 LC-CDR 1-3 aa: 135 aa: 157 No modification HV: D71T M87L; N93S aa: 4, 6, 8 nt: 1 nt: 43 nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-80 LC-CDR 1-3 aa: 135 aa: 158 No modification HV: W72R M87L; N93S aa: 4, 6, 8 nt: 1 nt: 45 nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-81 LC-CDR 1-3 aa: 135 aa: 159 No modification HV: Q82E M87L; N93S aa: 4, 6, 8 nt: 1 nt: 47 nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-82 LC-CDR 1-3 aa: 135 aa: 160 No modification HV: T87R M87L; N93S aa: 4, 6, 8 nt: 1 nt: 49 nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-83 LC-CDR 1-3 aa: 135 aa: 161 No modification HV: D113E M87L; N93S aa: 4, 6, 8 nt: 1 nt: 51 nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 54 nt: 11, 13, 53 MS-84 LC-CDR 1-3 aa: 162 aa: 163 KV: H9L, HV: T87R M87L; N93S aa: 4, 6, 8 nt: 55 nt: 57 KV: S12P, nt: 3, 5, 7 KV: S18P, HC-CDR 1-3 KV: R47Q, aa: 12, 14, 16 KV: T85A, nt: 11, 13, 15 KV: T90V MS-85 LC-CDR 1-3 aa: 164 aa: 136 KV: F2I, No modification M87L; N93S aa: 4, 6, 8 nt: 59 nt: 9 KV: D73G, nt: 3, 5, 7 KV: K74T HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-86 LC-CDR 1-3 aa: 135 aa: 165 No modification HV: P25S, M87L; N93S aa: 4, 6, 8 nt: 1 nt: 61 HV: N27Y, nt: 3, 5, 7 HV: L29F HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-87 LC-CDR 1-3 aa: 135 aa: 166 No modification HV: D71T, M87L; N93S aa: 4, 6, 8 nt: 1 nt: 63 HV: W72R nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-88 LC-CDR 1-3 aa: 135 aa: 167 No modification HV: P25S, M87L; N93S aa: 4, 6, 8 nt: 1 nt: 65 HV: N27Y, nt: 3, 5, 7 HV: L29F, HC-CDR 1-3 HV: D71T, aa: 12, 14, 16 HV: W72R nt: 11, 13, 15 MS-89 LC-CDR 1-3 aa: 168 aa: 169 KV: H9L HV: N27Y, M87L; N93S aa: 4, 6, 8 nt: 67 nt: 69 HV: D71T nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-90 LC-CDR 1-3 aa: 170 aa: 171 KV: H9L HV: P25S, M87L; N93S aa: 4, 6, 8 nt: 71 nt: 73 HV: N27Y, nt: 3, 5, 7 HV: L29F HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-91 LC-CDR 1-3 aa: 172 aa: 173 KV: H9L, HV: P25S, M87L; N93S aa: 4, 6, 8 nt: 75 nt: 77 KV: K74T HV: N27Y nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-92 LC-CDR 1-3 aa: 174 aa: 175 KV: F2I HV: Q46E, M87L; N93S aa: 4, 6, 8 nt: 79 nt: 81 HV: W72R, nt: 3, 5, 7 HV: T87R HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 .sup.1Residues 1-57 of the nucleotide sequence for the heavy and light chain variable domains encode signal peptides, which are not a part of the mature sequence for the heavy and light chain variable domains indicated by the sequence identifiers in this table. IgG1 LC: light chain sequence modification IgG1 HC: heavy chain sequence modification
TABLE-US-00002 TABLE 2 Round-2 variants. Light Chain (LC)-CDR 1-3 Light Chain Heavy Chain Amino acid (aa) SEQ ID NO: Variable Domain Variable Domain Nucleotide (nt) SEQ ID NO: Amino acid (aa) Amino acid (aa) IgG1 Light Chain IgG1 Heavy Chain Fc Domain Heavy Chain (HC)-CDR 1-3 SEQ ID NO: SEQ ID NO: Modification Modification Modification Molecule Amino acid (aa) SEQ ID NO: Nucleotide (nt) Nucleotide (nt) (Relative to (Relative to (Relative to Set Nucleotide (nt) SEQ ID NO: SEQ ID NO: SEQ ID NO: SEQ ID NO: 135) SEQ ID NO: 136) SEQ ID NO: 137) MS-93 LC-CDR 1-3 aa: 176 aa: 136 KV: F2I, KV: H9L No modification M87L; N93S aa: 4, 6, 8 nt: 83.sup.2 nt: 9 nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-94 LC-CDR 1-3 aa: 177 aa: 136 KV: F2I, KV: S18P No modification M87L; N93S aa: 4, 6, 8 nt: 85 nt: 9 nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-95 LC-CDR 1-3 aa: 178 aa: 136 KV: F2I, KV: D73G No modification M87L; N93S aa: 4, 6, 8 nt: 87 nt: 9 nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-96 LC-CDR 1-3 aa: 179 aa: 136 KV: F2I, KV: T85A No modification M87L; N93S aa: 4, 6, 8 nt: 89 nt: 9 nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-97 LC-CDR 1-3 aa: 180 aa: 136 KV: H9L, KV: S18P No modification M87L; N93S aa: 4, 6, 8 nt: 91 nt: 9 nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-98 LC-CDR 1-3 aa: 181 aa: 136 KV: H9L, KV: D73G No modification M87L; N93S aa: 4, 6, 8 nt: 93 nt: 9 nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-99 LC-CDR 1-3 aa: 182 aa: 136 KV: H9L, KV: T85A No modification M87L; N93S aa: 4, 6, 8 nt: 95 nt: 9 nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-100 LC-CDR 1-3 aa: 183 aa: 136 KV: S18P, KV: D73G No modification M87L; N93S aa: 4, 6, 8 nt: 97 nt: 9 nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-101 LC-CDR 1-3 aa: 184 aa: 136 KV: S18P, KV: T85A No modification M87L; N93S aa: 4, 6, 8 nt: 99 nt: 9 nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-102 LC-CDR 1-3 aa: 185 aa: 136 KV: D73G, KV: T85A No modification M87L; N93S aa: 4, 6, 8 nt: 101 nt: 9 nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-103 LC-CDR 1-3 aa: 186 aa: 136 KV: H9L, KV: S18P No modification M87L; N93S aa: 4, 6, 8 nt: 103 nt: 9 nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-104 LC-CDR 1-3 aa: 187 aa: 136 KV: F2I, KV: H9L, No modification M87L; N93S aa: 4, 6, 8 nt: 105 nt: 9 KV: D73G nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-105 LC-CDR 1-3 aa: 188 aa: 136 KV: F2I, KV: H9L, No modification M87L; N93S aa: 4, 6, 8 nt: 107 nt: 9 KV: T85A nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-106 LC-CDR 1-3 aa: 189 aa: 136 KV: F2I, KV: S18P, No modification M87L; N93S aa: 4, 6, 8 nt: 109 nt: 9 KV: D73G nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-107 LC-CDR 1-3 aa: 190 aa: 136 KV: F2I, KV: S18P, No modification M87L; N93S aa: 4, 6, 8 nt: 111 nt: 9 KV: T85A nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-108 LC-CDR 1-3 aa: 191 aa: 136 KV: F2I, KV: D73G, No modification M87L; N93S aa: 4, 6, 8 nt: 113 nt: 9 KV: T85A nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-109 LC-CDR 1-3 aa: 192 aa: 136 KV: H9L, KV: S18P, No modification M87L; N93S aa: 4, 6, 8 nt: 115 nt: 9 KV: D73G nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-110 LC-CDR 1-3 aa: 193 aa: 136 KV: H9L, KV: S18P, No modification M87L; N93S aa: 4, 6, 8 nt: 117 nt: 9 KV: T85A nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-111 LC-CDR 1-3 aa: 194 aa: 136 KV: H9L, KV: D73G, No modification M87L; N93S aa: 4, 6, 8 nt: 119 nt: 9 KV: T85A nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-112 LC-CDR 1-3 aa: 195 aa: 136 KV: S18P, KV: D73G, No modification M87L; N93S aa: 4, 6, 8 nt: 121 nt: 9 KV: T85A nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-113 LC-CDR 1-3 aa: 196 aa: 136 KV: F2I, KV: H9L, No modification M87L; N93S aa: 4, 6, 8 nt: 123 nt: 9 KV: S18P, KV: D73G nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-114 LC-CDR 1-3 aa: 197 aa: 136 KV: F2I, KV: H9L, No modification M87L; N93S aa: 4, 6, 8 nt: 125 nt: 9 KV: S18P, KV: T85A nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-115 LC-CDR 1-3 aa: 198 aa: 136 KV: F2I, KV: H9L, No modification M87L; N93S aa: 4, 6, 8 nt: 127 nt: 9 KV: D73G, KV: T85A nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-116 LC-CDR 1-3 aa: 199 aa: 136 KV: F2I, KV: S18P, No modification M87L; N93S aa: 4, 6, 8 nt: 129 nt: 9 KV: D73G, KV: T85A nt: 3, 5, 7 HC-CDR 1-3 aa: 12, 14, 16 nt: 11, 13, 15 MS-117 LC-CDR 1-3 aa: 200 aa: 136 KV: H9L, KV: S18P, No modification M87L; N93S aa: 4, 6, 8 nt: 131 nt: 9 KV: D73G, KV: T85A nt: 3, 5, 7 HC-CDR 1-3 aa: 12,14,16 nt: 11, 13, 15 MS-118 LC-CDR 1-3 aa: 201 aa: 136 KV: F2I, KV: H9L, No modification M87L; N93S aa: 4, 6, 8 nt: 133 nt: 9 KV: S18P, KV: D73G, nt: 3, 5, 7 KV: T85A HC-CDR 1-3 aa: 12,14,16 nt: 11, 13, 15 .sup.2Residues 1-57 of the nucleotide sequence for the heavy and light chain variable domains encode signal peptides, which are not a part of the mature sequence for the heavy and light chain variable domains indicated by the sequence identifiers in this table. IgG1 LC: light chain sequence modification IgG1 HC: heavy chain sequence modification
III. Biophysical Properties of the PGDM1400 Antibody Variants
[0250] PGDM1400 variant antibodies and antigen-binding fragments thereof that are produced by the optimization program described herein exhibit one or more of the following biophysical characteristics: increased low-pH stability; increased thermal stability; increased solubility; reduced aggregation; and increased intramolecular and thermodynamic stability, such as chemical stability, as determined by chemical unfolding. These biophysical attributes have been shown to be linked to improved manufacturability and storage stability.
Solubility
[0251] The PGDM1400 variant antibodies or fragments thereof described herein exhibit improved solubility, e.g., relative to the parental PGDM1400 antibody. The featured PGDM1400 variant antibodies or fragments thereof described herein exhibit solubility of at least about 1 mg/ml (e.g., about 0.1 mg/ml, 0.2 mg/ml, 0.3 mg/ml, 0.4 mg/ml, 0.5 mg/ml, 0.6 mg/ml, 0.7 mg/ml, 0.8 mg/ml, 0.9 mg/ml, 1 mg/ml, 1.5 mg/ml, 2.0 mg/ml, 2.5 mg/ml, 3.0 mg/ml, 3.5 mg/ml, 4.0 mg/ml, 4.5 mg/ml, 5.0 mg/ml, 5.5 mg/ml, 6.0 mg/ml, 6.5 mg/ml, 7.0 mg/ml, 7.5 mg/ml, 8.0 mg/ml, 8.5 mg/ml, 9.0 mg/ml, 9.5 mg/ml, or 10.0 mg/ml) in a solution containing about 6-10% PEG 10,000 (e.g., about 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7.0%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8.0%, 8.1%, 8.2%, 8.3%, 8.4%, 8.5%, 8.6%, 8.7%, 8.8%, 8.9%, 9.0%, 9.1%, 9.2%, 9.3%, 9.4%, 9.5%, 9.6%, 9.7%, 9.8%, 9.9%, or 10.0% PEG 10,000). In particular, at least 1 mg/ml of the antibody or fragment thereof is soluble in a solution with a concentration of 9.4% PEG 10,000. Improved solubility of the PGDM1400 variant antibodies and fragments thereof, relative to the native PGDM1400 antibody, increases efficient production (e.g., higher production titer) of the antibodies by minimizing the amounts of antibodies lost through precipitation (e.g., aggregation).
Thermal Stability
[0252] The PGDM1400 variant antibodies or fragments thereof described herein exhibit high thermal stability, e.g., relative to the parental PGDM1400 antibody. The PGDM1400 variant antibodies and fragments thereof described herein exhibit reduced degradation or resistance to degradation upon exposure to a wide range of temperature variations (e.g., thermal ramping at temperatures of between about 20-95° C.). Specifically, the PGDM1400 variant antibodies and fragments thereof described herein exhibit reduced degradation or resistance to degradation upon exposure to about 68° C. and/or about 69.2° C. Thermal stability of the PGDM1400 variant antibodies or fragments thereof described herein ensure their stability and sustainability when exposed to extreme non-physiologic conditions, such as conditions during manufacture or production of the antibodies. The improved thermal stability of the PGDM1400 variant antibodies or fragments thereof described herein contributes to their improved manufacturability. Improved thermal stability of the PGDM1400 variant antibodies or fragments thereof described herein also contributes to improved storage stability (e.g., stability when stored at a temperature of about −30° C. to about 25° C. (e.g., about −30° C., −25° C., −20° C., −15° C., −10° C., −5° C., 0° C., 5° C., 10° C., 15° C., 20° C., 25° C., 30° C., or 35° C.) over about 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 1 year, 2 years, 3 years, 4 years, 5 years, or more), making them more suitable for extended storage and subsequent therapeutic application.
Chemical Stability
[0253] The PGDM1400 variant antibodies or fragments thereof described herein exhibit increased chemical stability, e.g., relative to the parental PGDM1400 antibody. The featured PGDM1400 variant antibodies and fragments thereof exhibit chemical stability, as determined by chemical unfolding (e.g., as tested by guanidine hydrochloride (GuHCl) or urea concentrations, preferably by GuHCl concentrations). For example, the PGDM1400 variant antibodies described herein exhibit increased chemical stability at a final concentration of the antibody or fragment thereof of about 0.01-5.0 mg/ml (e.g., about 0.02 mg/ml, 0.03 mg/ml, 0.04 mg/ml, 0.05 mg/ml, 0.06 mg/ml, 0.07 mg/ml, 0.08 mg/ml, 0.09 mg/ml, 1.0 mg/ml, 1.5 mg/ml, 2.0 mg/ml, 2.5 mg/ml, 3.0 mg/ml, 3.5 mg/ml, 4.0 mg/ml, 4.5 mg/ml, or 5.0 mg/ml, for example at a final concentration of about 0.05 mg/ml) in the presence of GuHCl (e.g., a concentration of GuHCL of greater than about 0.001 M to about 6 M GuHCL), relative to the parental PGDM1400 antibody. In specific embodiments, the PGDM1400 variant antibody or fragment thereof (e.g., at a concentration of about 0.05 mg/ml) may exhibit reduced chemical unfolding in the presence of about 2.0 M or greater GuHCL (e.g., greater than 2.5 M, greater than 3.0 M, greater than 3.5 M, greater than 4.0 M, greater than 4.5 M, greater than 5.0 M, or greater than 5.5 M) GuHCl. For example, the PGDM1400 variant antibody or fragment thereof at a final concentration of about 0.05 mg/ml may exhibit reduced chemical unfolding relative to the parental PGDM1400 antibody (e.g., an equilibrium denaturation point) at a GuHCl concentration of about 2.0-2.5 M. The improved chemical stability of the PGDM1400 variant antibodies or fragments thereof described herein indicates that the PGDM1400 variant antibodies and fragments thereof exhibit improved stability and sustainability under various conditions, such as those during manufacture or production of the antibodies or fragments thereof. The chemical stability of the PGDM1400 variant antibodies or fragments thereof described herein thus contributes to the improved manufacturability of the same.
Low-pH Stability
[0254] The PGDM1400 variant antibodies or fragments thereof described herein exhibit improved stability at low pH, e.g., relative to the parental PGDM1400 antibody. The featured PGDM1400 variant antibodies and fragments thereof exhibit improved stability (e.g., reduced aggregation) when exposed to low pH, such as a pH less than about pH 5.0 (e.g., less than pH 4.6, less than pH 4.3, less than pH 4.0, less than pH 3.6, less than pH 3.3, or at pH 3.0 (e.g., at pH 3.3)). In preferred embodiments the featured PGDM1400 variant antibodies or fragments thereof exhibit improved stability at about pH 3.3, e.g., relative to the parental PGDM1400 antibody. The stability of the PGDM1400 variant antibodies or fragments thereof at low pH is measured in terms of reduced aggregation or resistance to aggregation upon exposure to the low pH conditions (for e.g., as assessed following neutralization to a higher pH). The featured PGDM1400 variant antibodies or fragments thereof do not aggregate or exhibit reduced aggregation (e.g., high molecular weight species) upon neutralization from low pH exposure, which in preferred embodiments is about pH 3.3. The improved low-pH stability of the PGDM1400 variant antibodies or fragments thereof described herein ensures their stability and sustainability when exposed to low pH or acidic conditions, e.g., during manufacture or production of the antibodies and fragments thereof. Low-pH stability of the PGDM1400 variant antibodies or fragments thereof described herein thus contributes to the improved manufacturability of the same.
Reduced Aggregation
[0255] The PGDM1400 variant antibodies or fragments thereof described herein exhibit reduced aggregation (e.g., reduced aggregation when exposed to low pH, solubilizing or chaotropic chemicals, and/or increased temperatures), e.g., relative to the parental PGDM1400 antibody. Aggregation can be evaluated by monitoring monomer content and/or oligomer content over time (e.g., over days, weeks, months, or years). The featured PGDM1400 variant antibodies and fragments thereof exhibit reduced aggregation (e.g., reduced levels of aggregation following low-pH (e.g., pH 3.3) incubation), as evaluated by monomer and/or oligomer content (e.g., monomer content more than about 60% (e.g., more than about 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, or 97%), and/or oligomer content less than about 10% (e.g., less than about 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.4%, or 0.3%)). Reduced aggregation of the PGDM1400 variant antibodies or fragments thereof described herein ensure their stability and sustainability when exposed to chemicals, low pH conditions, and extreme temperatures (e.g., large temperature variations or increased temperatures, e.g., in range of about −30° C. to about 35° C.) during manufacture or production of the antibodies or fragments thereof. Reduced aggregation of the PGDM1400 variant antibodies or fragments thereof described herein thus contributes to improved manufacturability of the same. Reduced aggregation of the PGDM1400 variant antibodies or fragments thereof described herein also ensures their stability during storage (e.g., storage for over about 2 days, over about 3 days, over about 4 days, over about 5 days, over about 6 days, over about 1 week, over about 2 weeks, over about 3 weeks, over about 1 month, over about 2 months, over about 3 months, over about 4 months, over about 5 months, over about 6 months, over about 7 months, over about 8 months, over about 9 months, over about 10 months, over about 11 months, over about 1 year, over about 2 years, over about 3 years, over about 4 years, over about 5 years, or more, at a temperature of about −30° C. to about 35° C. (e.g., about −30° C., −25° C., −20° C., −15° C., −10° C., −5° C., 0° C., 5° C., 10° C., 15° C., 20° C., 25° C., 30° C., or 35° C.)). Storage stability of the PGDM1400 variant antibodies or fragments thereof also ensures longer shelf life, retention of efficacy and safer therapeutic application of the same. With improved manufacturability and storage stability, the PGDM1400 variant antibodies or antigen-binding fragments thereof, featured herein, exhibit improved characteristics relative to the native PGDM1400 antibody.
Pharmacokinetics and Binding Affinity
[0256] The PGDM1400 variant antibodies or antigen-binding fragments thereof described herein exhibit a half-life of at least about 1 hour (e.g., at least about 1 hour, 2 hour, 3 hour, 4 hour, 5 hour, 6 hour, 7 hour, 8 hour, 9 hour, 10 hour, 11 hour, 12 hour, 13 hour, 14 hour 15 hour, 16 hour, 17 hour, 18 hour, 19 hour, 20 hour, 21 hour, 22 hour, 23 hour, 1 day, 2 day, 3 day, 4 day, 5 day, 6 day, 7 day, 8 day, 9 day, 10 day, 11 day, 12 day, 13 day, 14 day, 15 day, 16 day, 17 day, 18 day, 19 day, 20 day, 21 day, 22 day, 23 day, 24 day, 25 day, 26 day, 27 day, 28 day, or more) in vitro or in vivo (e.g., following administration to a subject (e.g., a human)). For example, the PGDM1400 variant antibodies or antigen-binding fragments thereof described herein may exhibit a half-life of at least about 1 hour in vivo (e.g., in a fluid, such as blood) following administration (e.g., intravenous administration) to a subject (e.g., a human).
[0257] The PGDM1400 variant antibodies or antigen-binding fragments thereof described herein may bind to a parental PGDM1400 anti-idiotype (ID) antibody. The PGDM1400 variant antibodies or antigen-binding fragments thereof described herein may exhibit the same affinity (e.g., binding affinity) for the parental PGDM1400 anti-ID antibody as the parental PGDM1400 antibody or have an affinity (e.g., binding affinity) for the parental PGDM1400 anti-ID antibody that is about ±10% of the affinity exhibited by the parental PGDM1400 antibody.
IV. Production of the PGDM1400 Antibody Variants
[0258] The PGDM1400 antibody variant or antigen-binding fragment thereof described herein may be in the form of a single-chain polypeptide, such as a scFv fragment. Single chain polypeptides may alternatively contain one or more CDRs described herein covalently bound to one another using conventional bond-forming techniques known in the art, for instance, by an amide bond, a thioether bond, a carbon-carbon bond, or by a linker, such as a peptide linker or a linker formed by nucleophilic substitution of a multi-valent electrophile (e.g., a bis(bromomethyl) arene derivative, such as a bis(bromomethyl)benzene or bis(bromomethyl)pyridine) described herein or known in the art.
[0259] Single-chain polypeptides can be produced by a variety of recombinant and synthetic techniques, such as by recombinant gene expression or solid-phase peptide synthesis procedures described herein or known in the art. For instance, one of skill in the art can design polynucleotides encoding, e.g., two or more CDRs operably linked to one another in frame so as to produce a continuous, single-chain peptide containing these CDRs. Optionally, the CDRs may be separated by a spacer, such as by a framework region (e.g., a framework sequence described herein or a framework region of a germline consensus sequence of a human antibody) or a flexible linker, such as a poly-glycine or glycine/serine linker described herein or known in the art. When produced by chemical synthesis methods, native chemical ligation can optionally be used as a strategy for the synthesis of long peptides (e.g., greater than 50 amino acids). Native chemical ligation protocols are known in the art and have been described, e.g., by Dawson et al. (Science, 266:776-779, 1994); incorporated herein by reference. A detailed description of techniques for the production of single-chain polypeptides, full-length antibodies, and antibody fragments is provided in the sections that follow.
[0260] The PGDM1400 antibody variant or antigen-binding fragment thereof described herein can be prepared by any of a variety of established techniques. For instance, an antibody or antigen-binding fragment thereof described herein can be prepared by recombinant expression of immunoglobulin light and heavy chain genes in a host cell. To express an antibody recombinantly, a host cell can be transfected with one or more recombinant expression vectors carrying DNA fragments encoding the immunoglobulin light and heavy chains of the antibody such that the light and heavy chains are expressed in the host cell and, optionally, secreted into the medium in which the host cells are cultured, from which medium the antibodies can be recovered. Standard recombinant DNA methodologies are used to obtain antibody heavy and light chain genes, incorporate these genes into recombinant expression vectors and introduce the vectors into host cells, such as those described in Molecular Cloning; A Laboratory Manual, Second Edition (Sambrook, Fritsch and Maniatis (eds), Cold Spring Harbor, N.Y., 1989), Current Protocols in Molecular Biology (Ausubel et al., eds., Greene Publishing Associates, 1989), and in U.S. Pat. No. 4,816,397; the disclosures of each of which are incorporated herein by reference.
Expression Vectors
[0261] Some methods for producing a PGDM1400 antibody variant or antigen-binding fragment thereof described herein involve expression in mammalian cells, although recombinant proteins can also be produced using insect cells, yeast, bacteria, or other cells under the control of appropriate promoters. Mammalian expression vectors may include non-transcribed elements such as an origin of replication, a suitable promoter and enhancer, and other 5′ or 3′ flanking non-transcribed sequences, and 5′ or 3′ non-translated sequences such as necessary ribosome binding sites, a polyadenylation site, splice donor and acceptor sites, and termination sequences. DNA sequences derived from the SV40 viral genome, for example, SV40 origin, early promoter, enhancer, splice, and polyadenylation sites may be used to provide the other genetic elements required for expression of a heterologous DNA sequence. Appropriate cloning and expression vectors for use with bacterial, fungal, yeast, and mammalian cellular hosts are described in Green & Sambrook, Molecular Cloning: A Laboratory Manual (Fourth Edition), Cold Spring Harbor Laboratory Press 2012.
[0262] Various mammalian cell culture systems can be employed to express and manufacture recombinant protein. Examples of mammalian expression systems include CHO cells, COS cells, HEK293, HeLA and BHK cell lines. Processes of culturing host cell for production of protein therapeutics are described in Zhou and Kantardjieff (Eds.), Mammalian Cell Cultures for Biologics Manufacturing (Advances in Biochemical Engineering/Biotechnology), Springer 2014.
[0263] Viral genomes also provide a rich source of vectors that can be used for the efficient delivery of exogenous genes into the genome of a cell (e.g., a eukaryotic or prokaryotic cell). Viral genomes are particularly useful vectors for gene delivery because the polynucleotides contained within such genomes are typically incorporated into the genome of a target cell by generalized or specialized transduction. These processes occur as part of the natural viral replication cycle, and do not require added proteins or reagents in order to induce gene integration. Examples of viral vectors include a retrovirus, adenovirus (e.g., Ad5, Ad26, Ad34, Ad35, and Ad48), parvovirus (e.g., adeno-associated viruses), coronavirus, negative strand RNA viruses such as orthomyxovirus (e.g., influenza virus), rhabdovirus (e.g., rabies and vesicular stomatitis virus), paramyxovirus (e.g., measles and Sendai), positive strand RNA viruses, such as picornavirus and alphavirus, and double stranded DNA viruses including adenovirus, herpesvirus (e.g., Herpes Simplex virus types 1 and 2, Epstein-Barr virus, cytomegalovirus), and poxvirus (e.g., vaccinia, modified vaccinia Ankara (MVA), fowlpox and canarypox). Other viruses useful for delivering polynucleotides encoding antibody light and heavy chains or antibody fragments described herein include Norwalk virus, togavirus, flavivirus, reoviruses, papovavirus, hepadnavirus, and hepatitis virus, for example. Examples of retroviruses include: avian leukosis-sarcoma, mammalian C-type, B-type viruses, D-type viruses, HTLV-BLV group, lentivirus, spumavirus (Coffin, J. M., Retroviridae: The viruses and their replication, In Fundamental Virology, Third Edition, B. N. Fields, et al., Eds., Lippincott-Raven Publishers, Philadelphia, 1996). Other examples include murine leukemia viruses, murine sarcoma viruses, mouse mammary tumor virus, bovine leukemia virus, feline leukemia virus, feline sarcoma virus, avian leukemia virus, human T cell leukemia virus, baboon endogenous virus, Gibbon ape leukemia virus, Mason Pfizer monkey virus, simian immunodeficiency virus, simian sarcoma virus, Rous sarcoma virus and lentiviruses. Other examples of vectors are described, for example, in McVey et al., (U.S. Pat. No. 5,801,030); the disclosures of each of which are incorporated herein by reference.
Genome Editing Techniques
[0264] In addition to viral vectors, a variety of additional methods have been developed for the incorporation of genes, e.g., those encoding antibody light and heavy chains, single-chain polypeptides, single-chain variable fragments (scFvs), tandem scFvs, Fab domains, F(ab′).sub.2 domains, diabodies, and triabodies, among others, into the genomes of target cells for polypeptide expression. One such method that can be used for incorporating polynucleotides encoding antibody variants, or antigen-binding fragments thereof (e.g., single-chain polypeptides, antibodies, antigen-binding fragments thereof, or constructs), into prokaryotic or eukaryotic cells includes transposons. Transposons are polynucleotides that encode transposase enzymes and contain a polynucleotide sequence or gene of interest flanked by excision sites at the 5′ and 3′ positions. Once a transposon has been delivered into a cell, expression of the transposase gene commences and results in active enzymes that cleave the gene of interest from the transposon. This activity is mediated by the site-specific recognition of transposon excision sites by the transposase. In some embodiments, these excision sites may be terminal repeats or inverted terminal repeats. Once excised from the transposon, the gene of interest can be integrated into the genome of a prokaryotic or eukaryotic cell by transposase-catalyzed cleavage of similar excision sites that exist within nuclear genome of the cell. This allows the gene encoding the antibody variant described in the invention or fragment or domain thereof to be inserted into the cleaved nuclear DNA at the excision sites, and subsequent ligation of the phosphodiester bonds that join the gene of interest to the DNA of the prokaryotic or eukaryotic cell genome completes the incorporation process. In some embodiments, the transposon may be a retrotransposon, such that the gene encoding the antibody is first transcribed to an RNA product and then reverse-transcribed to DNA before incorporation in the prokaryotic or eukaryotic cell genome. Exemplary transposon systems include the piggybac transposon (described in detail in WO 2010/085699) and the sleeping beauty transposon (described in detail in US20050112764); the disclosures of each of which are incorporated herein by reference.
[0265] Another useful method for the integration of nucleic acid molecules encoding the antibody or antigen-binding fragments thereof (e.g., single-chain polypeptides, antibodies, or antigen-binding fragments thereof) into the genome of a prokaryotic or eukaryotic cell is the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas system, which is a system that originally evolved as an adaptive defense mechanism in bacteria and archaea against infection by viruses. The CRISPR/Cas system consists of palindromic repeat sequences within plasmid DNA and an associated Cas9 nuclease. This ensemble of DNA and protein directs site specific DNA cleavage of a target sequence by first incorporating foreign DNA into CRISPR loci. Polynucleotides containing these foreign sequences and the repeat-spacer elements of the CRISPR locus are in turn transcribed in a host cell to create a guide RNA, which can subsequently anneal to a target sequence and localize the Cas9 nuclease to this site. In this manner, highly site-specific cas9-mediated DNA cleavage can be engendered in a foreign polynucleotide because the interaction that brings cas9 within close proximity of the target DNA molecule is governed by RNA:DNA hybridization. As a result, one can theoretically design a CRISPR/Cas system to cleave any target DNA molecule of interest. This technique has been exploited in order to edit eukaryotic genomes (Hwang et al., Nat. Biotech., 31:227-229, 2013) and can be used as an efficient means of site-specifically editing eukaryotic or prokaryotic genomes in order to cleave DNA prior to the incorporation of a polynucleotide encoding a PGDM1400 antibody variant (e.g., single-chain polypeptides, antibodies, or antigen-binding fragments thereof) described herein. The use of CRISPR/Cas to modulate gene expression has been described in U.S. Pat. No. 8,697,359, the disclosure of which is incorporated herein by reference.
[0266] Alternative methods for site-specifically cleaving genomic DNA prior to the incorporation of a polynucleotide encoding an antibody or antibody fragment described herein include the use of zinc finger nucleases and transcription activator-like effector nucleases (TALENs). Unlike the CRISPR/Cas system, these enzymes do not contain a guiding polynucleotide to localize to a specific target sequence. Target specificity is instead controlled by DNA binding domains within these enzymes. Zinc finger nucleases and TALENs for use in genome editing applications are described in Urnov et al. (Nat. Rev. Genet., 11:636-646, 2010); and in Joung et al., (Nat. Rev. Mol. Cell. Bio. 14:49-55, 2013); incorporated herein by reference. Additional genome editing techniques that can be used to incorporate polynucleotides encoding antibodies described herein into the genome of a prokaryotic or eukaryotic cell include the use of ARCUS™ meganucleases that can be rationally designed so as to site-specifically cleave genomic DNA. The use of these enzymes for the incorporation of polynucleotides encoding antibodies (e.g., antibodies, antigen-binding fragments thereof, or constructs) described herein into the genome of a prokaryotic or eukaryotic cell is particularly advantageous in view of the structure-activity relationships that have been established for such enzymes. Single-chain meganucleases can thus be modified at certain amino acid positions in order to create nucleases that selectively cleave DNA at desired locations. These single-chain nucleases have been described extensively, e.g., in U.S. Pat. Nos. 8,021,867 and 8,445,251; the disclosures of each of which are incorporated herein by reference.
Polynucleotide Sequence Elements
[0267] To express antibodies (e.g., single-chain polypeptides, antibodies, antigen-binding fragments thereof, or constructs) described herein, polynucleotides encoding partial or full-length light and heavy chains, e.g., polynucleotides that encode one or more, or all, of the CDR sequences of a PGDM1400 antibody variant or antigen-binding fragment thereof described herein can be inserted into an expression vector such that the nucleic acid molecules encoding the PGDM1400 antibody variant sequences are operatively linked to transcriptional and translational control sequences. The expression vector and expression control sequences are chosen to be compatible with the expression host cell used. Polynucleotides encoding the light chain and the heavy chain domains of a PGDM1400 antibody variant or fragment thereof described herein can be inserted into separate vectors, or, optionally, both polynucleotides can be incorporated into the same expression vector using established techniques described herein or known in the art.
[0268] In addition to polynucleotides encoding the heavy and light chains of a PGDM1400 antibody variant (or a polynucleotide encoding a single-chain polypeptide, an antibody fragment, such as a scFv molecule, or a construct described herein), the recombinant expression vectors described herein may carry regulatory sequences that control the expression of the antibody chain polynucleotides in a host cell. The design of the expression vector, including the selection of regulatory sequences, may depend on such factors as the choice of the host cell to be transformed or the level of expression of protein desired. For instance, suitable regulatory sequences for mammalian host cell expression include viral elements that direct high levels of protein expression in mammalian cells, such as promoters and/or enhancers derived from cytomegalovirus (CMV) (such as the CMV promoter/enhancer), Simian Virus 40 (SV40) (such as the SV40 promoter/enhancer), adenovirus, (e.g., the adenovirus major late promoter (AdMLP)) and polyoma. Viral regulatory elements, and sequences thereof, are described in detail, for instance, in U.S. Pat. Nos. 5,168,062, 4,510,245, and 4,968,615, the disclosures of each of which are incorporated herein by reference.
[0269] In addition to the antibody heavy and light chain polynucleotides and regulatory sequences, the recombinant expression vectors described herein can carry additional sequences, such as sequences that regulate replication of the vector in host cells (e.g., origins of replication) and selectable marker genes. A selectable marker gene facilitates selection of host cells into which the vector has been introduced (see e.g., U.S. Pat. Nos. 4,399,216, 4,634,665 and 5,179,017). For example, typically the selectable marker gene confers resistance to cytotoxic drugs, such as G418, puromycin, blasticidin, hygromycin or methotrexate, to a host cell into which the vector has been introduced. Suitable selectable marker genes include the dihydrofolate reductase (DHFR) gene (for use in DHFR″ host cells with methotrexate selection/amplification) and the neo gene (for G418 selection). In order to express the light and heavy chain domains of a PGDM1400 antibody or antigen-binding fragment thereof, the expression vector(s) containing polynucleotides encoding the heavy and light chain domains can be transfected into a host cell by standard techniques.
V. Antiretroviral Agents (ARVs) for Use in Combination with PGDM1400 Variant Antibodies
[0270] In certain instances, a PGDM1400 variant antibody or fragment thereof featured herein may be used in combination with one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) antiretroviral agents (ARVs), such as, without limitation, any one or more ARVs set forth in Table 3 below.
TABLE-US-00003 TABLE 3 Antiretroviral Agents Generic Name (Brand Name) Class efavirenz, emtricitabine and tenofovir Multi-class disoproxil fumarate (Atripla) emtricitabine, rilpivirine, and tenofovir Multi-class disoproxil fumarate (Complera) elvitegravir, cobicistat, emtricitabine, Multi-class tenofovir disoproxil fumarate (Stribild) lamivudine and zidovudine (Combivir) NRTI emtricitabine, FTC (Emtriva) NRTI lamivudine, 3TC (Epivir) NRTI abacavir and lamivudine (Ebzicom) NRTI zalcitabine, dideoxycytidine, ddC (Hivid) NRTI zidovudine, azidothymidine, AZT, ZDV NRTI (Retrovir) abacavir, zidovudine, and lamivudine NRTI (Trizivir) tenofovir disoproxil fumarate and NRTI emtricitabine (Truvada) enteric coated didanosine, ddI EC (Videx NRTI EC) didanosine, dideoxyinosine, ddI (Videx) NRTI tenofovir disoproxil fumarate, TDF (Viread) NRTI stavudine, d4T (Zerit) NRTI abacavir sulfate, ABC (Ziagen) NRTI Rilpivirine (Edurant) NNRTI Etravirine (Intelence) NNRTI delavirdine, DLV (Rescriptor) NNRTI efavirenz, EFV (Sustiva) NNRTI nevirapine, NVP (Viramune) NNRTI nevirapine, NVP (Viramune XR) NNRTI amprenavir, APV (Agenerase) PI tipranavir, TPV (Aptivus) PI indinavir, IDV (Crixivan) PI saquinavir (Fortovase) PI saquinavir mesylate, SQV (Invirase) PI lopinavir and ritonavir, LPV/RTV (Kaletra) PI Fosamprenavir Calcium, FOS-APV (Lexiva) PI ritonavir, RTV (Norvir) PI Darunavir (Prezista) PI atazanavir sulfate, ATV (Reyataz) PI nelfinavir mesylate, NFV (Viracept) PI enfuvirtide, T-20 (Fuzeon) Fusion Inhibitor maraviroc (Selzentry) Entry Inhibitor - CCR5 co-receptor antagonist raltegravir (Isentress) HIV integrase strand transfer inhibitors dolutegravir (Tivicay) HIV integrase strand transfer inhibitors
[0271] One or more of the above ARVs may be used (e.g., administered to a subject in need thereof) in combination with a PGDM1400 variant antibody or fragment thereof featured herein, and, optionally, one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) HIV-specific broadly neutralizing antibody (bnAb), such as a CD4bs-specific antibody (e.g., 3BNC117 or VRC07-523), an N332 glycan-dependent antibody (e.g., PGT121, or a variant thereof), and/or a V2-specific antibody (e.g., CAP256-VRC26 and/or the parental PGDM1400). One or more of the above ARVs may be administered to a subject (e.g., a human), either alone, or in combination with the bnAb, prior to, concurrently with, and/or subsequent to administration of the antibody (e.g., a PGDM1400 variant antibody or fragment thereof) featured herein.
VI. Immunomodulators for Use in Combination with PGDM1400 Variant Antibodies
[0272] A PGDM1400 variant antibody or fragment thereof featured herein may be used in combination with one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) immunomodulators, such as, without limitation, any one or more immunomodulators set forth in Table 4 below.
TABLE-US-00004 TABLE 4 Exemplary Immunomodulators Drug Name AS-101 Bropirimine Acemannan CL246,738 EL10 FP-21399 Gamma Interferon Granulocyte Macrophage Colony Stimulating Factor HIV Core Particle Immunostimulant Interleukin-2 (IL-2) Immune Globulin Intravenous (human) IMREG-1 IMREG-2 Imuthiol Diethyl Dithio Carbamate Alpha-2 Interferon Methionine-Enkephalin MTP-PE Muramyl-Tripeptide Granulocyte Colony Stimulating Factor Remune rCD4-IgG hybrids Recombinant Soluble Human CD4 SK&F106528 Soluble T4 Thymopentin Tumor Necrosis Factor Infliximab
[0273] One or more of the above immunomodulators may be used (e.g., administered to a subject in need thereof) in combination with a PGDM1400 variant antibody or fragment thereof featured herein, and, optionally, one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) HIV-specific bnAb, such as a CD4bs-specific antibody (e.g., 3BNC117 or VRC07-523), an N332 glycan-dependent antibody (e.g., PGT121, or a variant thereof), a V2-specific antibody (e.g., CAP256-VRC26 and/or the parental PGDM1400), and/or one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) ARVs. One or more of the above immunomodulators may be administered to a subject (e.g., a human), either alone, or in combination with the bnAb and/or the ARV, prior to, concurrently with, and/or subsequent to administration of the PGDM1400 variant antibody or antigen-binding fragment thereof featured herein.
VII. Reservoir Activators for Use in Combination with PGDM1400 Variant Antibodies
[0274] A PGDM1400 variant antibody or fragment thereof featured herein may be used in combination with one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) reservoir activators, such as, without limitation, any one or more reservoir activators described by Spivak and Planelles (Annu Rev Med, 69:421-436, 2018), Stoszko et al (EBioMedicine, 3:108-121, 2016), and Delagreverie et al (Open Forum Infectious Diseases, DOI: 10.1093/ofid/ofw189); incorporated herein by reference. Examples of reservoir activators that may be used in combination with a PGDM1400 variant antibody or fragment thereof featured herein are set forth in Table 5 below.
TABLE-US-00005 TABLE 5 Exemplary reservoir activators Class of agents Agents PKC agonists (i) Phorbol esters, including phorbol 12-myristate 13-acetate (PMA), prostratin and 12-deoxyphorbol 13-phenylacetate (DPP); (ii) Macrocyclic lactones including bryostatin-1 and analogs (iii) Diterpenes, including ingenol compounds Cytokines and IL-7, IFN-α, IL-15 superagonist ALT-803 (IL-15N72D + chemokines IL-15RαSu/Fc fusion protein) Toll-like (i) TLR 1/2 agonists, including Pam3CSK4 receptor (ii) TLR3 agonists, including Poly-ICLC (TLR) agonists (iii) TLR5 agonists, including flagellin (iv) TLR7 agonists, including GS-9620 (v) TLR9 agonists, including MGN1703 and CpG7909 Immune Anti-PD-1 monoclonal antibodies, anti-PD-1 ligand checkpoint (PD-L1) monoclonal antibodies, anti-CTLA-4 inhibitors monoclonal antibodies HDAC romidepsin, vorinostat, belinostat, LAQ824, inhibitors panobinostat, entinostat, CI994, mocetinostat Small (i) Disulfiram molecules (ii) Benzotriazole derivatives, including 3-Hydroxy-1,2,3-benzotriazin-4((3H)-one (HO-DHBt) (iii) SMAC mimetics (iv) BRG-Brahma Associated Factor (BAF) inhibitors, including caffeic acid phenethyl ester and pyrimethamine
[0275] One or more of the above reservoir activators may be used (e.g., administered to a subject in need thereof) in combination with a PGDM1400 variant antibody or fragment thereof featured herein, and, optionally, one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) HIV-specific bnAb, such as a CD4bs-specific antibody (e.g., 3BNC117 or VRC07-523), an N332 glycan-dependent antibody (e.g., PGT121, or a variant thereof), a V2-specific antibody (e.g., CAP256-VRC26 and/or the parental PGDM1400), one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) ARVs, and/or one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) immunomodulators. One or more of the above reservoir activators may be administered to a subject (e.g., a human), either alone, or in combination with the bnAb, the ARV, and/or the immunomodulator, prior to, concurrently with, and/or subsequent to administration of the PGDM1400 variant antibody or fragment thereof featured herein.
VIII. Therapeutic Methods
[0276] The PGDM1400 variant antibodies or fragments thereof described herein can be administered to a subject in need thereof to treat or block HIV infection in the subject. In one or more methods described herein, the featured PGDM1400 variant antibody or fragment thereof can be administered, either alone, or in combination with one or more of a bnAb, ARV, reservoir activator and/or immunomodulator, to a subject (e.g., a human) in need thereof to cure HIV infection in the subject. In particular, featured are methods of treating a subject (e.g., a human) infected with HIV (e.g., HIV-1), in which the methods include administering to the subject one or more of the PGDM1400 variant antibodies or antigen-binding fragments thereof described hereinabove. These methods are supported by the findings that the PGDM1400 variant antibodies or fragments thereof described herein are capable of neutralizing pseudoviruses of HIV, such as SC422661.8, RHPA4259.7, Du172.17, BB1012-11.TC21, CNE52, 0260.v5.c36, 263-8, SC05.8C11.2344, X1193_c1, Ce1176_A3, AC10.0.29, and 6952.v1.c20.
[0277] Included are methods of blocking an HIV (e.g., HIV-1) infection in a subject (e.g., a human) at risk of HIV transmission by administering one or more of the PGDM1400 variant antibodies and/or antigen binding fragments thereof to the subject. For example, in one aspect, the subject may be a fetus of an HIV-infected pregnant female and the method includes administering to the HIV-infected pregnant female one or more of the PGDM1400 variant antibodies or antigen-binding fragments thereof described hereinabove, thereby blocking the HIV infection in the fetus. In other instances, the subject is a newborn having an HIV-infected mother, a subject at risk of HIV transmission following a needlestick injury, or a subject at risk of HIV transmission following a sexual exposure to one or more HIV-infected individuals. In instances when the subject is a fetus of an HIV-infected pregnant female, the HIV-infected pregnant female can be administered one or more of the PGDM1400 variant antibodies or antigen-binding fragments thereof described hereinabove following manifestation of one or more symptoms associated with pregnancy (e.g., a missed period, tender or swollen breasts, nausea with or without vomiting, increased urination, fatigue, and/or uncharacteristic food aversions or cravings), following a diagnosis of pregnancy, and/or in the third trimester of pregnancy, in order to block an HIV infection in the fetus.
[0278] In instances when the subject is a newborn having an HIV-infected mother, the newborn can be administered one or more of the PGDM1400 variant antibodies or antigen-binding fragments thereof described hereinabove peripartum and/or postpartum, for example, prior to, during, and/or following breastfeeding from the HIV-infected mother, in order to block an HIV infection in the newborn.
[0279] In instances when the subject is at risk of HIV transmission following a needlestick injury, the subject can be administered one or more of the PGDM1400 variant antibodies or antigen-binding fragments thereof described hereinabove less than 3 days following the needlestick injury, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 35, 40, 45, 50, 55, or 60 minutes, 2, 4, 6, 10, 15, or 24 hours, 1.5, 2, or 2.5 days following the needlestick injury, in order to block an HIV infection in the subject. Alternatively, or additionally, the subject can be administered one or more of the PGDM1400 variant antibodies or antigen-binding fragments thereof described hereinabove between 3 to 14 days following the needlestick injury, for example, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 days following the needlestick injury, in order to block an HIV infection in the subject.
[0280] In instances when the subject is at risk of HIV transmission following a sexual exposure to one or more HIV-infected individuals, the subject can be administered one or more of the PGDM1400 variant antibodies or antigen-binding fragments thereof described hereinabove less than 3 days following the sexual exposure, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 35, 40, 45, 50, 55, or 60 minutes, 2, 4, 6, 10, 15, or 24 hours, 1.5, 2, or 2.5 days following the sexual exposure, in order to block an HIV infection in the subject. Alternatively, or additionally, the subject can be administered one or more of the PGDM1400 variant antibodies or antigen-binding fragments thereof described hereinabove between 3 to 14 days following the sexual exposure, for example, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 days following the sexual exposure, in order to block an HIV infection in the subject.
[0281] In any of the methods of antibody therapy described above, the subject can have an undetectable plasma viral load, such as less than 3,500 RNA copies/ml (e.g., less than 2,000 RNA copies/ml, e.g., less than 400 RNA copies/ml, e.g., less than 50 RNA copies/ml, e.g., less than 1 RNA copy/ml), prior to commencement of antibody therapy. In such instances, the subject may already be on ARV. However, ARV alone, in contrast to the PGDM1400 variant antibodies or antigen-binding fragments thereof described hereinabove, is unable to reduce tissue reservoirs of the virus. Accordingly, the methods of the invention feature administration of one or more of the PGDM1400 variant antibodies or antigen-binding fragments thereof described hereinabove, alone or in combination with one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) ARV, and/or one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) HIV-specific bnAb (such as a CD4bs-specific antibody (e.g., 3BNC117 or VRC07-523), an N332 glycan-dependent antibody (e.g., PGT121, or a variant thereof), and/or a V2-specific antibody (e.g., CAP256-VRC26 and/or the parental PGDM1400)), as described in detail below, to treat a subject (e.g., a human) infected with HIV (e.g., HIV-1) or block an HIV infection in a subject at risk of HIV transmission, based, at least in part, on the finding that the PGDM1400 variant antibodies or fragments thereof described hereinabove are capable of neutralizing pseudoviruses of HIV, such as RHPA4259.7, Du172.17, CNE52, 0260.v5.c36, SC05.8C11.2344, Ce1176_A3, SC422661.8, BB1012-11.TC21, 263-8, X1193_c1, AC10.0.29, and 6952.v1.c20. Preferably, the subject either maintains or achieves an undetectable plasma viral load for at least about 2 months (e.g., at least about 3, 4, 5, 6, 7, 8, 9, 10, or 11 months, or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 years) following administration of the PGDM1400 variant antibodies or fragments thereof described hereinabove. The reduction in plasma viral load may be in the absence of an ART, e.g., for a period of at least about 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, 12 years, 13 years, 14 years, 15 years, 16 years, 17 years, 18 years, 19 years, 20 years, or more after administration of the PGDM1400 variant antibody or antigen-binding fragment thereof.
[0282] In any of the methods described above, further administration of an immunomodulator (e.g., an agent, such as a protein or peptide, which is capable of increasing, inducing, or extending an immune response, e.g., a cell-mediated immune response and/or a humoral immune response, when administered to a subject, such as a human, e.g., a human infected with HIV or at risk of an HIV infection or transmission) is contemplated. For example, one or more immunomodulators (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more immunomodulators) can be administered in conjunction with, e.g., prior to, concurrently with, subsequent to, or within the context of a treatment regimen that includes administration of a PGDM1400 variant antibody or fragment thereof described hereinabove.
[0283] In any of the methods described above, further administration of a reservoir activator (e.g., one or more reservoir activators selected from Table 5) is contemplated. For example, one or more reservoir activators (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more reservoir activators) can be administered in conjunction with, e.g., prior to, concurrently with, subsequent to, or within the context of a treatment regimen that includes administration of a PGDM1400 variant antibody or fragment thereof described hereinabove.
[0284] In any of the methods described above, administration of one or more of the PGDM1400 variant antibodies or antigen-binding fragments thereof described hereinabove, alone or in combination with one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) HIV-specific bnAb (such as a CD4bs-specific antibody (e.g., 3BNC117 or VRC07-523), an N332 glycan-dependent antibody (e.g., PGT121, or a variant thereof), and/or a V2-specific antibody (e.g., CAP256-VRC26 and/or the parental PGDM1400)), one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) ARVs, one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) reservoir activators, and/or one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) immunomodulators may: (i) reduce proviral DNA to below about 1,000 DNA copies/10.sup.6 cells (e.g., below about 900 DNA copies/10.sup.6 cells, below about 800 DNA copies/10.sup.6 cells, below about 700 DNA copies/10.sup.6 cells, below about 600 DNA copies/10.sup.6 cells, below about 500 DNA copies/10.sup.6 cells, below about 400 DNA copies/10.sup.6 cells, below about 300 DNA copies/10.sup.6 cells, below about 200 DNA copies/10.sup.6 cells, below about 100 DNA copies/10.sup.6 cells, below about 90 DNA copies/10.sup.6 cells, below about 80 DNA copies/10.sup.6 cells, below about 70 DNA copies/10.sup.6 cells, below about 60 DNA copies/10.sup.6 cells, below about 50 DNA copies/10.sup.6 cells, below about 40 DNA copies/10.sup.6 cells, below about 30 DNA copies/10.sup.6 cells, below about 20 DNA copies/10.sup.6 cells, below about 10 DNA copies/10.sup.6 cells, below about 9 DNA copies/10.sup.6 cells, below about 8 DNA copies/10.sup.6 cells, below about 7 DNA copies/10.sup.6 cells, below about 6 DNA copies/10.sup.6 cells, below about 5 DNA copies/10.sup.6 cells, below about 4 DNA copies/10.sup.6 cells, below about 3 DNA copies/10.sup.6 cells, below about 2 DNA copies/10.sup.6 cells, below about 1 DNA copy/10.sup.6 cells, or to an undetectable level) in a tissue (e.g., lymph node tissue, gastrointestinal tissue, peripheral blood) of the subject relative to an untreated control; (ii) increase HIV-specific cell-mediated immune response and/or humoral immune response in the subject relative to an untreated control; (iii) decrease viral replication in the subject relative to an untreated control; and/or (iv) reduce the plasma viral load to less than about 3,500 RNA copies/ml (e.g., less than about 3,000 RNA copies/ml, less than about 2,500 RNA copies/ml, less than about 2,000 RNA copies/ml, less than about 1,500 RNA copies/ml, less than about 1,000 RNA copies/ml, less than about 550 RNA copies/ml, less than about 500 RNA copies/ml, less than about 450 RNA copies/ml, less than about 400 RNA copies/ml, less than about 350 RNA copies/ml, less than about 300 RNA copies/ml, less than about 250 RNA copies/ml, less than about 200 RNA copies/ml, less than about 150 RNA copies/ml, less than about 100 RNA copies/ml, less than about 50 RNA copies/ml, less than about 40 RNA copies/ml, less than about 30 RNA copies/ml, less than about 20 RNA copies/ml, less than about 10 RNA copies/ml, less than about 9 RNA copies/ml, less than about 8 RNA copies/ml, less than about 7 RNA copies/ml, less than about 6 RNA copies/ml, less than about 5 RNA copies/ml, less than about 4 RNA copies/ml, less than about 3 RNA copies/ml, less than about 2 RNA copies/ml, less than about 1 RNA copy/ml, or to an undetectable level) relative to an untreated control. In some instances, following administration of one or more of the PGDM1400 variant antibodies or antigen-binding fragments thereof described hereinabove, alone or in combination with one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) HIV-specific bnAb (such as a CD4bs-specific antibody (e.g., 3BNC117 or VRC07-523), an N332 glycan-dependent antibody (e.g., PGT121, or a variant thereof), and/or a V2-specific antibody (e.g., CAP256-VRC26 and/or the parental PGDM1400)), one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) ARVs, one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) reservoir activators, and/or one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) immunomodulators, the subject has an undetectable plasma viral load for at least about 2 months (e.g., at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 1 year, at least about 2 years, at least about 3 years, at least about 4 years, at least about 5 years, at least about 6 years, at least about 7 years, at least about 8 years, at least about 9 years, at least about 10 years, at least about 11 years, at least about 12 years, at least about 13 years, at least about 14 years, at least about 15 years, at least about 16 years, at least about 17 years, at least about 18 years, at least about 19 years, at least about 20 years, or more).
[0285] As described below in more detail, in any of the methods described above, the HIV therapy (e.g., HIV-1 therapy) may be concluded following administration of at least one dose (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more doses) of the PGDM1400 variant antibody or antigen-binding fragment thereof described hereinabove, alone or in combination with one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) HIV-specific bnAb (such as a CD4bs-specific antibody (e.g., 3BNC117 or VRC07-523), an N332 glycan-dependent antibody (e.g., PGT121, or a variant thereof), and/or a V2-specific antibody (e.g., CAP256-VRC26 and/or the parental PGDM1400)), one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) ARVs, one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) reservoir activators, and/or one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) immunomodulators, following a duration of time post-therapy (e.g., at least about two months or longer). The subject (e.g., a human infected with HIV or at risk of HIV transmission) can be monitored post-therapy to confirm that they exhibit and/or maintain virologic control in the absence of any intervening therapies, which, optionally, can be determined based upon measurements made from a biological sample of the subject (e.g., a measurement of proviral DNA level in a tissue and/or plasma viral load). If the subject exhibits and/or maintains virologic control during this post-therapy period, the subject may be taken off one or more, or all, HIV therapies indefinitely or until such time as the subject begins to exhibit loss of virologic control.
IX. Methods of Administration and Dosage
[0286] For any of the methods describe above, the one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) PGDM1400 variant antibodies or antigen-binding fragments thereof described hereinabove can be formulated, dosed, and administered in a fashion consistent with good medical practice. Antibody therapy may be performed alone or in conjunction with another therapy (e.g., ARV therapy or administration of a reservoir activator), and may be provided at home, the doctor's office, a clinic, a hospital's outpatient department, or a hospital. Antibody therapy optionally begins at a hospital so that the doctor can observe the therapy's effects closely and make any adjustments that are needed, or it may begin on an outpatient basis.
[0287] The dosage administered can be selected based on the subject to be treated (e.g., the age, body weight, capacity of the immune system, and general health of the subject being treated), the form of administration (e.g., as a solid or liquid), the manner of administration (e.g., by injection, inhalation, dry powder propellant), and the cells targeted (e.g., mucosal cells, epithelial cells, such as blood vessel epithelial cells, nasal epithelial cells, or pulmonary epithelial cells). Additionally, pharmacogenomic (the effect of genotype on the pharmacokinetic, pharmacodynamic, or efficacy profile of a therapeutic) information about a particular subject may affect the dosage used. Antibody therapy of the invention is preferably administered in an amount that provides a sufficient level of one or more of the PGDM1400 variant antibodies or antigen-binding fragments thereof to yield a therapeutic effect in the subject without undue adverse physiological effects caused by treatment.
[0288] An PGDM1400 variant antibody or antigen-binding fragment thereof described hereinabove can be administered to a subject (e.g., a human infected with HIV and/or at risk of HIV transmission) intramuscularly, intravenously, intradermally, intraarterially, intraperitoneally, intralesionally, intracranially, intraarticularly, intraprostatically, intrapleurally, intratracheally, intranasally, intravitreally, intravaginally, intrarectally, topically, intratumorally, peritoneally, subcutaneously, subconjunctival, intravesicularly, mucosally, intrapericardially, intraumbilically, intraocularally, orally, topically, locally, by inhalation, by injection, by infusion, by continuous infusion, by localized perfusion bathing target cells directly, by catheter, by lavage, in cremes, or in lipid compositions, in accord with known methods. For example, the PGDM1400 variant antibody or antigen-binding fragment thereof described hereinabove can be administered by infusion, such as by continuous infusion (e.g., intravenously). Alternatively, it is envisioned that the PGDM1400 variant antibody or antigen-binding fragment thereof described hereinabove may be delivered by gene therapy.
[0289] For any of the methods described above, a single dose of a PGDM1400 variant antibody or antigen-binding fragment thereof described hereinabove can be administered to the subject. The single dose may be of a single PGDM1400 variant antibody or antigen-binding fragment thereof described hereinabove or of more than one antibody (i.e., an antibody cocktail including multiple antibodies or antigen-binding fragments thereof described hereinabove). In some instances, HIV therapy (e.g., HIV-1 therapy) may be concluded following the administration of the single dose of the PGDM1400 variant antibody or fragment thereof described hereinabove. In some instances, the single dose may be administered along with one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) ARVs, such as one or more of the ARVs listed in Table 3 above, wherein the ARV is administered concurrently with, prior to (e.g., about 1 year, 9 months, 6 months, 3 months, 1 month, 3 weeks, 2 weeks, 1 week, 5 days, 3 days, 1 day, 18 hours, 12 hours, 6 hours, or 1 hour prior to), and/or subsequent to (e.g., about 1 year, 9 months, 6 months, 3 months, 1 month, 3 weeks, 2 weeks, 1 week, 5 days, 3 days, 1 day, 18 hours, 12 hours, 6 hours, or 1 hour subsequent to) the single dose of the PGDM1400 variant antibody or fragment thereof described hereinabove. Accordingly, HIV therapy can, in some instances, be concluded following the administration of the ARV subsequent to the single dose of the PGDM1400 variant antibody or fragment thereof described hereinabove.
[0290] Alternatively, or additionally, the single dose may be administered along with a one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) HIV-specific bnAb (such as a CD4bs-specific antibody (e.g., 3BNC117 or VRC07-523), an N332 glycan-dependent antibody (e.g., PGT121, or a variant thereof), and/or a V2-specific antibody (e.g., CAP256-VRC26 and/or the parental PGDM1400)), wherein the HIV-specific bnAb is administered concurrently with, prior to (e.g., about 1 year, 9 months, 6 months, 3 months, 1 month, 3 weeks, 2 weeks, 1 week, 5 days, 3 days, 1 day, 18 hours, 12 hours, 6 hours, or 1 hour prior to), and/or subsequent to (e.g., about 1 year, 9 months, 6 months, 3 months, 1 month, 3 weeks, 2 weeks, 1 week, 5 days, 3 days, 1 day, 18 hours, 12 hours, 6 hours, or 1 hour subsequent to) the single dose of the PGDM1400 variant antibody or fragment thereof described hereinabove, alone, or in combination with one or more ARV. Accordingly, HIV therapy can, in some instances, be concluded following the administration of the HIV-specific bnAb (e.g., 3BNC117, VRC07-523, PGT121 or variant thereof, CAP256-VRC26, or the parental PGDM1400) subsequent to the single dose of the PGDM1400 variant antibody or fragment thereof described hereinabove.
[0291] Alternatively, or additionally, the single dose may be administered along with a one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) immunomodulators (e.g., one or more immunomodulators selected from Table 4), wherein the immunomodulator is administered concurrently with, prior to (e.g., about 1 year, 9 months, 6 months, 3 months, 1 month, 3 weeks, 2 weeks, 1 week, 5 days, 3 days, 1 day, 18 hours, 12 hours, 6 hours, or 1 hour prior to), and/or subsequent to (e.g., about 1 year, 9 months, 6 months, 3 months, 1 month, 3 weeks, 2 weeks, 1 week, 5 days, 3 days, 1 day, 18 hours, 12 hours, 6 hours, or 1 hour subsequent to) the single dose of the PGDM1400 variant antibody or fragment thereof described hereinabove, alone, or in combination with one or more ARV, and/or HIV-specific bnAb (e.g., 3BNC117, VRC07-523, PGT121 or variant thereof, CAP256-VRC26, or the parental PGDM1400). Accordingly, HIV therapy can, in some instances, be concluded following the administration of the immunomodulator subsequent to the single dose of the PGDM1400 variant antibody or fragment thereof described hereinabove.
[0292] Alternatively, or additionally, the single dose may be administered along with a one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) reservoir activators (e.g., one or more reservoir activators selected from Table 5), wherein the reservoir activator is administered concurrently with, prior to (e.g., about 1 year, 9 months, 6 months, 3 months, 1 month, 3 weeks, 2 weeks, 1 week, 5 days, 3 days, 1 day, 18 hours, 12 hours, 6 hours, or 1 hour prior to), and/or subsequent to (e.g., about 1 year, 9 months, 6 months, 3 months, 1 month, 3 weeks, 2 weeks, 1 week, 5 days, 3 days, 1 day, 18 hours, 12 hours, 6 hours, or 1 hour subsequent to) the single dose of the PGDM1400 variant antibody or fragment thereof described hereinabove, alone, or in combination with one or more ARV, HIV-specific bnAb (e.g., 3BNC117, VRC07-523, PGT121 or variant thereof, CAP256-VRC26, or the parental PGDM1400), and/or immunomodulators. Accordingly, HIV therapy can, in some instances, be concluded following the administration of the reservoir activators subsequent to the single dose of the PGDM1400 variant antibody or fragment thereof described hereinabove.
[0293] In other instances, the method includes administering a first regimen including one or more doses (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more doses) of the PGDM1400 variant antibody or fragment thereof described hereinabove and a second regimen including one or more doses (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more doses) of the PGDM1400 variant antibody or fragment thereof described hereinabove, wherein the second regimen is administered at least about 2 months (e.g., at least about 3, 4, 5, 6, 7, 8, 9, 10, or 11 months, or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 years) after the first regimen. The duration of time between the first and second regimens is preferably a longer duration of time than necessary for viral rebound to occur in a subject (e.g., a human) infected with HIV (e.g., HIV-1) under current standard of care (e.g., ART), which is approximately two months. Thus, the second regimen of the PGDM1400 variant antibody or fragment thereof described hereinabove can be considered a maintenance dose, and in some instances, HIV therapy may be concluded following the administration of the second regimen of the PGDM1400 variant antibody or fragment thereof described hereinabove. In some instances, the method can further include administering one or more (e.g., 1, 2, 3, 4, or 5 or more) ARV, such as one or more of the ARVs listed in Table 3 above, wherein the ARV is administered concurrently with, prior to, and/or subsequent to the first regimen and/or the second regimen of the PGDM1400 variant antibody or fragment thereof described hereinabove. Accordingly, HIV therapy can, in some instances, be concluded following the administration of the ARV subsequent to the second regimen of the PGDM1400 variant antibody or fragment thereof described hereinabove. Alternatively, or additionally, the first and second regimens may be administered along with a HIV-specific bnAb, such as CD4bs-specific antibodies (e.g., 3BNC117 or VRC07-523), an N332 glycan-dependent antibody (e.g., PGT121, or a variant thereof), and/or a V2-specific antibody (e.g., CAP256-VRC26 and/or the parental PGDM1400). Accordingly, HIV therapy can, in some instances, be concluded following the administration of the HIV-specific bnAb (e.g., 3BNC117, VRC07-523, PGT121 or variant thereof, CAP256-VRC26, or the parental PGDM1400) subsequent to second regimen of the PGDM1400 variant antibody or fragment thereof described hereinabove. Alternatively, or additionally, the first and second regimens may be administered along with an immunomodulator, such as one or more of the immunomodulators listed in Table 4 above. Accordingly, HIV therapy can, in some instances, be concluded following the administration of the immunomodulator subsequent to second regimen of the PGDM1400 variant antibody or fragment thereof described hereinabove. Alternatively, or additionally, the first and second regimens may be administered along with a reservoir activator, such as one or more of the reservoir activators listed in Table 5 above. Accordingly, HIV therapy can, in some instances, be concluded following the administration of the reservoir activator subsequent to second regimen of the PGDM1400 variant antibody or fragment thereof described hereinabove.
[0294] For any of the methods described above, a PGDM1400 variant antibody or fragment thereof described hereinabove can be administered to the subject in a unit dose form or as a dose per mass or weight of the subject from about 0.01 mg/kg to about 100 mg/kg (e.g., about 0.01-0.1 mg/kg, e.g., 0.02 mg/kg, 0.03 mg/kg, 0.04 mg/kg, 0.05 mg/kg, 0.06 mg/kg, 0.07 mg/kg, 0.08 mg/kg, 0.09 mg/kg, 0.1 mg/kg, e.g., 0.1-1 mg/kg, e.g., 0.2 mg/kg, 0.3 mg/kg, 0.4 mg/kg, 0.5 mg/kg, 0.6 mg/kg, 0.7 mg/kg, 0.8 mg/kg, 0.9 mg/kg, 1 mg/kg, e.g., 1-10 mg/kg, e.g., 1 mg/kg, 2 mg/kg, 3 mg/kg, 4 mg/kg, 5 mg/kg, 6 mg/kg, 7 mg/kg, 8 mg/kg, 9 mg/kg, 10 mg/kg, e.g., 10-100 mg/kg, e.g., 20 mg/kg, 30 mg/kg, 40 mg/kg, 50 mg/kg, 60 mg/kg, 70 mg/kg, 80 mg/kg, 90 mg/kg, 100 mg/kg). The PGDM1400 variant antibody or fragment thereof described hereinabove can be administered to the subject at a dose of about 0.01-100 mg/kg (e.g., about 0.02-100 mg/kg, 0.03-100 mg/kg, 0.04-mg/kg, 0.05-100 mg/kg, 0.06-100 mg/kg, 0.07-100 mg/kg, 0.08-100 mg/kg, 0.09-100 mg/kg, 0.1-90 mg/kg, 0.1-80 mg/kg, 0.1-70 mg/kg, 0.1-60 mg/kg, 0.1-50 mg/kg, 0.5-50 mg/kg, 0.5-40 mg/kg, 0.5-30 mg/kg, 0.5-20 mg/kg, 0.5-10 mg/kg, 0.5-5 mg/kg, or 0.5-1 mg/kg) per mass or weight of the subject. For any of the methods described above, about 0.01-5000 mg (e.g., about 0.01-4500 mg, 0.01-4000 mg, 0.01-3500 mg, 0.01-3000 mg, 0.01-2500 mg, 0.01-2000 mg, 0.01-1500 mg, 0.01-1000 mg, 0.05-1000 mg, 0.1-1000 mg, 0.1-500 mg, 0.5-500 mg, 0.5-450 mg, 0.5-400 mg, 0.5-350 mg, 0.5-300 mg, 0.5-250 mg, 0.5-200 mg, 0.5-150 mg, 0.5-100 mg, 0.5-50 mg, 0.5-45 mg, 0.5-40 mg, 0.5-35 mg, 0.5-30 mg, 0.5-25 mg, 0.5-20 mg, 0.5-15 mg, 0.5-10 mg, or 1-10 mg) of the PGDM1400 variant antibody or fragment thereof described hereinabove can be administered to the subject.
[0295] A PGDM1400 variant antibody or fragment thereof described hereinabove may be administered to the subject two or more times, such as one or more times hourly, daily (e.g., once daily for up to six days), weekly, every two weeks, every three weeks, every four weeks, monthly, every two months, every three months, every six months, or every year. The method may further include administering a second dose of the PGDM1400 variant antibody or fragment thereof described hereinabove to the subject about one week, two weeks, three weeks, four weeks, or five weeks after administration of a first dose of the PGDM1400 variant antibody or fragment thereof described hereinabove. The method may also include administering more than two doses (e.g., three, four, five, six, seven, eight, nine, ten, or more doses) of the PGDM1400 variant antibody or fragment thereof to the subject. Administration of a PGDM1400 variant antibody or fragment thereof described hereinabove can be repeated at such a frequency for a certain period of time, followed by a period without treatment. Such repeated administrations can occur over a course of therapy lasting a specified length of time (e.g., at least about 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, or more).
[0296] In some of the methods of the invention, HIV (e.g., HIV-1) therapy is concluded following a determination that the proviral DNA level in tissue of the subject (as assessed, e.g., by biopsy) is reduced to an undetectable level. The method can result in a reduction of proviral DNA level in tissue of the subject relative to an amount of proviral DNA level in tissue of the subject before the administration of the PGDM1400 variant antibody or fragment thereof described hereinabove, or relative to an untreated control. For example, the proviral DNA level in tissue (e.g., lymph node tissue, gastrointestinal tissue, and/or peripheral blood) may be reduced to an undetectable level, such as below about 1,000 DNA copies/10.sup.6 cells (e.g., below about 100 DNA copies/10.sup.6 cells, e.g., below about 10 DNA copies/10.sup.6 cells, e.g., below about 1 DNA copy/10.sup.6 cells). Thus, a definitive end to HIV therapy can be determined based upon measurements made from a biological sample of the subject and/or time post-administration of the PGDM1400 variant antibody or fragment thereof described hereinabove.
[0297] According to any one of the methods of the invention described herein, a PGDM1400 variant antibody or fragment thereof described hereinabove can be administered as a pharmaceutical composition. The pharmaceutical composition has the antibody or antigen-binding fragment thereof alone, or in combination with one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) ARV (e.g., one or more ARVs selected from Table 3), one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) immunomodulators (e.g., one or more immunomodulators selected from Table 4), one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) reservoir activators (e.g., one or more reservoir activators selected from Table 5), and/or one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) HIV-specific bnAb (e.g., 3BNC117, VRC07-523, PGT121, CAP256-VRC26, or PGDM1400). The pharmaceutical composition has the antibody or antigen-binding fragment thereof in an amount of about 0.01-5000 mg (e.g., about 0.01-4000 mg, 0.01-3000 mg, 0.01-2000 mg, 0.01-1000 mg, 0.05-1000 mg, 0.05-500 mg, 0.05-400 mg, 0.05-300 mg, 0.05-200 mg, 0.05-100 mg, 0.1-100 mg, 0.1-90 mg, 0.1-80 mg, 0.1-70 mg, 0.1-60 mg, 0.1-50 mg, 0.1-40 mg, 0.1-30 mg, 0.1-20 mg, 0.1-10 mg, 0.1-9 mg, 0.1-8 mg, 0.1-7 mg, 0.1-6 mg, 0.1-5 mg, 0.1-4 mg, 0.1-3 mg, 0.1-2 mg, or 0.1-1 mg). The pharmaceutical composition with the antibody or antigen-binding fragment thereof may be formulated in a volume of about 1000 ml or less (e.g., about 950 ml or less, about 900 ml or less, about 850 ml or less, about 800 ml or less, about 750 ml or less, about 700 ml or less, about 650 ml or less, about 600 ml or less, about 550 ml or less, about 500 ml or less, about 450 ml or less, about 400 ml or less, about 350 ml or less, about 300 ml or less, about 250 ml or less, about 200 ml or less, about 150 ml or less, about 100 ml or less, about 50 ml or less, about 25 ml or less, about 20 ml or less, about 15 ml or less, about 10 ml or less, about 5 ml or less, about 1 ml or less, or about 0.1 ml or less). The pharmaceutical composition with the PGDM1400 variant antibody or antigen-binding fragment thereof may be formulated in a volume of about 900 ml, 850 ml, 800 ml, 750 ml, 700 ml, 650 ml, 600 ml, 550 ml, 500 ml, 450 ml, 400 ml, 350 ml, 300 ml, 250 ml, 200 ml, 150 ml, 100 ml, 50 ml, 25 ml, 20 ml, 15 ml, 10 ml, 9 ml, 8 ml, 7 ml, 6 ml, 5 ml, 4 ml, 3 ml, 2 ml, 1 ml, 0.5 ml, 0.1 ml, 0.05 ml, or 0.01 ml. The pharmaceutical composition with the PGDM1400 variant antibody or antigen-binding fragment thereof may be formulated in a volume of about 0.1-10 ml (e.g., about 0.1-9 ml, 0.1-8 ml, 0.1-7 ml, 0.1-6 ml, 0.1-5 ml, 0.1-4 ml, 0.1-3 ml, 0.1-2 ml, or 0.1-1 ml)
[0298] Methods of formulating pharmaceutical agents are known in the art, e.g., Niazi, Handbook of Pharmaceutical Manufacturing Formulations (Second Edition), CRC Press 2009, describes formulation development for liquid, sterile, compressed, semi-compressed and OTC forms. Transdermal and mucosal delivery, lymphatic system delivery, nanoparticles, controlled drug release systems, theranostics, protein and peptide drugs, and biologics delivery are described in Wang et al., Drug Delivery: Principles and Applications (Second Edition), Wiley 2016; formulation and delivery of peptide and protein agent is described, e.g., in Banga, Therapeutic Peptides and Proteins: Formulation, Processing, and Delivery Systems (Third Edition), CRC Press 2015. The pharmaceutical composition may be formulated to release the PGDM1400 variant antibody or fragment thereof described hereinabove immediately upon administration (e.g., targeted delivery) or at any predetermined time period after administration using controlled or extended release formulations. Administration of the pharmaceutical composition in controlled or extended release formulations is useful where the composition, either alone or in combination, has (i) a narrow therapeutic index (e.g., the difference between the plasma concentration leading to harmful side effects or toxic reactions and the plasma concentration leading to a therapeutic effect is small; generally, the therapeutic index, TI, is defined as the ratio of median lethal dose (LD.sub.50) to median effective dose (ED.sub.50)); (ii) a narrow absorption window at the site of release (e.g., the gastro-intestinal tract); or (iii) a short biological half-life, so that frequent dosing during a day is required in order to sustain a therapeutic level.
[0299] Many strategies can be pursued to obtain controlled or extended release in which the rate of release outweighs the rate of metabolism of the pharmaceutical composition. For example, controlled release can be obtained by the appropriate selection of formulation parameters and ingredients, including, e.g., appropriate controlled release compositions and coatings. Suitable formulations are known to those of skill in the art. Examples include single or multiple unit tablet or capsule compositions, oil solutions, suspensions, emulsions, microcapsules, microspheres, nanoparticles, patches, and liposomes.
[0300] The pharmaceutical compositions may be sterilized by conventional sterilization techniques, or may be sterile filtered. The resulting aqueous solutions may be packaged for use as is or lyophilized. The lyophilized preparation may be administered in powder form or combined with a sterile aqueous carrier prior to administration. The pH of the preparations typically will be between 3 and 11, more preferably between 5 and 9 or between 6 and 8, and most preferably between 7 and 8, such as 7 to 7.5. The resulting pharmaceutical compositions in solid form may, for example, be packaged in multiple single-dose units, each containing a fixed amount of a PGDM1400 variant antibody or fragment thereof described hereinabove, and, if desired, one or more immunomodulatory agents, reservoir activators, HIV-specific bnAbs (such as CD4bs-specific antibodies (e.g., 3BNC117 or VRC07-523), an N332 glycan-dependent antibody (e.g., PGT121, or a variant thereof), and/or a V2-specific antibody (e.g., CAP256-VRC26 and/or the parental PGDM1400)), and/or ARVs, such as in a sealed package of tablets or capsules, or in a suitable dry powder inhaler (DPI) capable of administering one or more doses.
[0301] The pharmaceutical compositions, including a PGDM1400 variant antibody or fragment thereof described hereinabove, can be prepared using standard methods known in the art by mixing the active ingredient (e.g., a PGDM1400 variant antibody or antigen-binding fragment thereof described hereinabove) having the desired degree of purity with, optionally, pharmaceutically acceptable carriers, excipients, or stabilizers (Remington's Pharmaceutical Sciences (20th edition), ed. A. Gennaro, 2000, Lippincott, Williams & Wilkins, Philadelphia, Pa.). Acceptable carriers, include saline, or buffers such as phosphate, citrate and other organic acids; antioxidants including ascorbic acid; low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone, amino acids such as glycine, glutamine, asparagines, arginine or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; salt-forming counterions such as sodium; and/or nonionic surfactants such as TWEEN™, PLURONICS™, or PEG.
[0302] A PGDM1400 variant antibody or fragment thereof described hereinabove can be administered in a pharmaceutical composition that includes one or more pharmaceutically acceptable carriers, excipients, or diluents. Examples of suitable carriers, excipients, or diluents include, e.g., saline, sterile water, polyalkylene glycols, oils of vegetable origin, hydrogenated napthalenes, suitable buffer, 1,3-butanediol, Ringer's solution and/or sodium chloride solution. Exemplary formulations for parenteral administration includes solutions prepared in water suitably mixed with a surfactant, e.g., hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, DMSO and mixtures thereof with or without alcohol, and in oils. Under ordinary conditions of storage and use, these preparations may contain a preservative to prevent the growth of microorganisms. Other exemplary carriers, excipients, or diluents are described in the Handbook of Pharmaceutical Excipients, 6th Edition, Rowe et al., Eds., Pharmaceutical Press (2009), hereby incorporated by reference in its entirety.
[0303] A pharmaceutical composition can be formulated to be compatible with its intended route of administration. Solutions or suspensions used for parenteral, intradermal, or subcutaneous application includes the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose. pH can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide. The parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.
[0304] Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, or phosphate buffered saline (PBS). In all cases, the composition must be sterile and should be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, and sodium chloride in the composition. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate and gelatin.
[0305] Sterile injectable solutions can be prepared by incorporating the PGDM1400 variant antibody or fragment thereof in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization. Dispersions can be prepared by incorporating a PGDM1400 variant antibody or antigen-binding fragment thereof into a sterile vehicle which contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation can be vacuum drying and freeze-drying which yields a powder of the PGDM1400 variant antibody or antigen-binding fragment thereof plus any additional desired ingredient from a previously sterile-filtered solution thereof.
[0306] Oral compositions include an inert diluent or an edible carrier. The composition can be enclosed in a gelatin capsule or compressed into a tablet. For the purpose of oral therapeutic administration, a PGDM1400 variant antibody or fragment thereof can be incorporated with excipients and used in the form of tablets, troches, or gelatin capsules. Oral compositions can also be prepared using a fluid carrier for use as a mouthwash, wherein the compound in the fluid carrier is applied orally and swished and expectorated or swallowed. Pharmaceutically compatible binding agents, and/or adjuvant materials can be included as part of the composition. The tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, or corn starch; a lubricant such as magnesium stearate; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring.
[0307] Systemic administration can also be by transmucosal or transdermal means. For transmucosal or transdermal administration, penetrants appropriate to the barrier to be permeated can be used in the formulation. Such penetrants are generally known, and include, for example, for transmucosal administration, detergents, bile salts, and fusidic acid derivatives. Transmucosal administration can be accomplished through the use of nasal sprays or suppositories. For transdermal administration, the antibody or antigen-binding fragment thereof may be formulated into ointments, salves, gels, or creams as generally known in the art.
[0308] A PGDM1400 variant antibody or fragment thereof can be prepared with carriers that will protect the compound against rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Methods for preparation of such formulations will be apparent to those skilled in the art. Liposomal suspensions can also be used as pharmaceutically acceptable carriers. These can be prepared according to methods known to those skilled in the art.
[0309] The pharmaceutical compositions can be included in a container, pack, or dispenser together with instructions for administration.
[0310] Optionally, but preferably, the formulation contains a pharmaceutically acceptable salt, preferably sodium chloride, and preferably at about physiological concentrations. Optionally, the formulations of the invention can contain a pharmaceutically acceptable preservative. In some embodiments the preservative concentration ranges from 0.1 to 2.0%, typically v/v. Suitable preservatives include those known in the pharmaceutical arts. Benzyl alcohol, phenol, m-cresol, methylparaben, and propylparaben are preferred preservatives. Optionally, the formulations of the invention include a pharmaceutically acceptable surfactant at a concentration of 0.005 to 0.02%.
X. Kits
[0311] Also featured herein are kits that include the aforementioned PGDM1400 antibody variant or antigen-binding fragment thereof, the polynucleotide encoding the PGDM1400 antibody variant or antigen-binding fragment thereof, the vector containing the polynucleotide, the host cell with the polynucleotide or the vector (e.g., a prokaryotic cell or a eukaryotic cell (e.g., a mammalian cell, such as a CHO or a HEK293 cell)), or the aforementioned composition (e.g., composition including the aforementioned PGDM1400 antibody variant or antigen-binding fragment thereof, the polynucleotide encoding the antibody or antigen-binding fragment thereof, the vector containing the polynucleotide, or the host cell with the polynucleotide or the vector (e.g., a prokaryotic cell or a eukaryotic cell (e.g., a mammalian cell, such as a CHO or a HEK293 cell)), and, e.g., a pharmaceutically-acceptable carrier, in a therapeutically effective amount for preventing or treating HIV infection (e.g., HIV-1 infection) in a subject (e.g., a human, such as a human infected with HIV). The kits can include instructions directing a clinician (e.g., a physician or nurse) in methods for administering the PGDM1400 antibody variant or antigen-binding fragment thereof, the polynucleotide, the vector, the host cell or the composition contained therein.
[0312] The kits may include multiple packages of single-dose pharmaceutical composition(s) containing an effective amount of a PGDM1400 antibody variant or antigen-binding fragment thereof, polynucleotide encoding the PGDM1400 antibody variant or antigen-binding fragment thereof, vector containing the polynucleotide, cell with the polynucleotide or composition featured herein. Optionally, instruments or devices necessary for administering the pharmaceutical composition(s) may be included in the kits. For instance, a kit of this invention may provide one or more pre-filled syringes containing an effective amount of the composition described herein (e.g., composition including one or more of the PGDM1400 antibody variant(s) or antigen-binding fragment(s) thereof, as described herein). Furthermore, the kits may also include additional components, such as instructions or schedules for administration of the composition to a patient infected with or at risk of being infected with HIV (e.g., HIV-1).
[0313] It will be apparent to those skilled in the art that various modifications and variations can be made in the compositions, methods, and kits of the invention without departing from the spirit or scope of the invention. Thus, it is intended that the invention cover the modifications and variations of this invention provided they come within the scope of the appended claims and their equivalents.
EXAMPLES
[0314] The present invention is illustrated by the following examples, which are in no way intended to be limiting of the invention.
Example 1. Generation of bNAbs
Materials and Methods
[0315] Antibody material was generated from transient expression of two suspension cell lines, Human Embryonic Kidney 293 (HEK293) and Chinese Hamster Ovary (CHO). The pTT5 mammalian expression vectors containing either a light chain (LC) or heavy chain (HC) coding region were co-transfected into HEK293 cells at a viable cell density (VCD) of 1×10.sup.6 cells/mL using polyethyleneimine (PEI) (Durocher et al., Nucleic Acids Res 30(2):E9, 2002), and then two-fold diluted with pre-warmed medium to ⅕ shake flask volume. Expression duration was 5-7 days at 37° C., 5% CO.sub.2, and 85% humidity at a shaking speed of 130 RPM with an orbit of 19 mm. All clarified supernatants were produced by pelleting the cells at 3000 g for 20 minutes followed by 0.22 μm filtration.
[0316] Antibodies were purified from the clarified supernatants using MABSELECT SURE™ protein A resin. A sodium phosphate, sodium chloride buffer system with an arginine wash and an acetate pH 3.5 elution was utilized. Protein A elutions were neutralized with tris, and buffer exchanged into 20 mM sodium phosphate, 150 mM NaCl, pH 7.4.
Size Exclusion High Performance Liquid Chromatography (SE-HPLC)
[0317] Size exclusion high performance liquid chromatography (SE-HPLC) was used to separate proteins based on differences in their hydrodynamic volumes. By this method, molecules with larger hydrodynamic protein volumes elute earlier than molecules with smaller volumes. Undiluted samples were loaded onto a Waters XBRIDGE® Protein BEH SEC 200A column (3.5 μm, 7.8×300 mm), separated isocratically with a running buffer (100 mM sodium phosphate and 250 mM sodium chloride, pH 6.8), and the eluent monitored by UV absorbance at 280 nm. Purity was determined by calculating the percentage of each separated component as compared to the total integrated area.
Differential Scanning Fluorimetry (DSF)
[0318] Differential scanning fluorimetry (DSF) is a high throughput technique that is used to estimate a protein's relative thermodynamic stability. Ranking of DSF results can be used as a tool to select candidates with more favorable stability properties. The DSF technique consists of measuring the fluorescence intensity of a hydrophobic probe at gradually increasing temperatures to determine the transition temperature and exposure of the hydrophobic regions of a protein. The measurements from this technique, reported as transition temperatures, correlate well with data obtained from differential scanning calorimetry (DSC). Thermal transition temperature(s) by DSF were measured according to the method of Feng et al. (J Pharm Sci 99: 1707-1720, 2010). Analysis was done in PBS buffer (20 mM sodium phosphate and 150 mM sodium chloride, pH 7.1) at a final protein concentration of 0.15 mg/ml and a final SYPRO® Orange concentration of 3×. Protein and SYPRO® Orange were mixed at 1:1 volumetric ratio in a 96-well PCR plate and analyzed using a Roche LIGHTCYCLER® 480 instrument equipped with Thermal Shift Analysis Software. Thermal curves were generated by heating the samples from 20-95° C. at a ramp rate of 4.4° C./s and 10 acquisitions per ° C. at Ex=465 nm Em=580 nm. Transition temperatures and shoulder scores were determined using the first derivative of the melting curve.
Thermal Hold Analysis
[0319] The stability of proteins at various temperatures was determined as follows. Samples are place in a 96-well Bio-Rad PCR plate and heated to various temperatures for 5 minutes using a Bio-Rad Thermal Cycler. After heating, samples were transferred to a 384-well Grenier clear plate. Protein precipitation was determined by reading the absorbance at 350 nm (A350) using a SPECTRAMAX® M5 plate reader.
Low-pH Stability
[0320] The stability of proteins at low pH was determined as follows. The pH of protein samples (1 mg/ml in 20 mM PBS) was lowered to approximately pH 3.3 using 2 M acetic acid. After a 30 minute incubation, samples were neutralized to approximately pH 5.0 using 2 M tris base. Samples were measured in duplicates for high molecular weight species using the SE-HPLC method. As a control, protein samples had the same volume of PBS added as the 2 M acetic acid and 2 M tris base, and measured for high molecular weight species.
Relative Solubility
[0321] Solubility was assessed according to the method of Torprani et al. (J Pharm Sci 105: 2319-2327, 2016). Analysis was done in PBS buffer (20 mM sodium phosphate and 150 mM sodium chloride, pH 7.1) and a final PEG 10,000 concentration of 7.9%. Protein at 1 mg/ml was diluted into the PEG solution at 1:4 ratio, and incubated in a 96-well 0.22 μm filter plate overnight at room temperature. After PEG incubation, samples were passed through the filter by centrifugation, and the remaining soluble protein was measured by a protein A titer assay.
Chemical Unfolding
[0322] Thirty-two guanidine hydrochloride (GuHCl) concentrations in PBS ranging from 0 to 6 M GND were prepared using a liquid handling robot. Protein samples (1 mg/ml in 20 mM PBS) were then transferred to each GuHCl concentration to achieve a final protein concentration of 0.05 mg/ml. After a 24 hour incubation, the samples were measured on a SPECTRAMAX® M5 plate reader (excitation: 280 nm, emission: 300-450 nm). The measured fluorescence intensity at 373 nm was corrected for scattering and stray light by subtraction of a small amount of the summed intensity measured between 300-320 nm (used as a surrogate for signal due to scattering), and then ratioed to the total intensity measured between 320-440 nm to correct for total intensity fluctuations. Then, the chemical unfolding curve was generated by plotting each corrected intensity against the GuHCl concentration. The inflection point of the curve was calculated and reported for each protein sample from this curve. Samples were measured in triplicate.
Neutralization Activity Assay
[0323] Neutralization titers of monoclonal antibodies (mAb) were determined using a luciferase-based assay in TZM.bl cells, according to the methods of Montefiori et al. (Methods Mol Biol 485: 395-405, 2009) and Sarzotti-Kelsoe et al. (J Immunol Methods 409: 131-146, 2014). Briefly, mAb samples at a primary concentration of 25 μg/ml with 5-fold serial dilutions were tested against a panel of 10 HIV-1 pseudoviruses that were selected for being PGDM1400 sensitive. Following incubation of antibody titers with HIV-1 pseudoviruses for 1 hour at 37° C., TZM.bl cells were added in growth media containing DEAE-dextran at a final concentration of 11 μg/ml. Assay plates were incubated for 48 hours at 37° C. and 5% CO.sub.2, and luciferase reporter gene expression was measured using BRIGHT-GLO™ luciferase reagent (Promega) and a VICTOR3™ luminometer (PerkinElmer). Neutralization titers (50% and 80% inhibitory concentrations, IC50 and IC80, respectively) were calculated as the mAb concentration at which relative light unit (RLU) was reduced by 50% or 80% compared to RLU in virus control wells after subtraction of background RLU in cell control wells. All assays were performed in a laboratory meeting GCLP standards.
Example 2. Development of Optimized PGDM1400 Variant Antibodies
[0324] A series of algorithms were applied to identify potentially destabilizing residues in the Fv region of the broadly neutralizing antibody, PGDM1400. These residues by themselves, or in combination, lead to instability at low pH, increased susceptibility to chemical degradation, or increased aggregation during production or long term storage. Based on this analysis, a series of variants were designed for maintaining potency while optimizing desired characteristics using combinatorial residue replacement techniques. The optimization process was broken up into different stages, the first being identification of single residues in the framework region that are potentially responsible for destabilization. Based on the analysis, a series of variants were produced by transient expression, each containing a single residue modification of the identified amino acids, or in a few variants, combinations of amino acids based on their proximity to each other (Round-1 variants, Table 1;
Example 3. Characterization of Round-1 PGDM1400 Variant Antibodies
[0325] Round-1 variants of PGDM1400 (Table 1) were produced by transient expression in HEK293 cells and purified by protein A chromatography. The antibodies were buffer exchanged into phosphate buffered saline and used for analysis. Assays used for analysis of the Round-1 variants included titer, size exclusion chromatography (SEC) to quantify high molecular weight (HMW) species and oligomers following purification (Table 6), DSF to characterize stability of the CH2 and Fab domains during thermal ramping, chemical unfolding by GuHCl for determining storage stability (Table 7), PEG solubility to interrogate protein-protein interaction (Table 8), and retention of neutralization capacity (Tables 9 and 10).
[0326] The monomer content of the variants ranged from a low of 88.2% to a high of 92%, where majority of the variation was due to dimer formation in the protein A purified material. DSF analysis showed that the Tm1 varied from 69.1 to 71.4° C., with a small number of the single variants (e.g., MS-66, MS-67, MS-70 and MS-75) possessing a Tm2. Weighted Shoulder Score (WSS) analysis, which provides a finer distinction between variants, with higher values being more desirable, showed that a subset of the single mutation variant with KV:F2I mutation (e.g., MS-66) had a 20 point increase in WSS over the parental molecule, MS-119. Other biophysical assays correlated to stability, including chemical unfolding that reports inflection point and ΔG of unfolding, and solubility in PEG solutions also showed increased values for variants with single point mutations. Incubation of the parental antibody and variants in the low pH solution followed by neutralization showed little change in the HMW values, indicating that the molecule was stable under low pH conditions.
[0327] The variants were also assayed for retention of neutralization activity. Tables 9 and 10 show neutralization activity of Round-1 variants against 12 pseudoviruses of HIV, which are representative of the broader set of viruses against which the parental PGDM1400 antibody is active. The PGDM1400 variant antibodies with more than 3-fold increase in the 1050 or 1080 values for a particular pseudovirus were considered inactive and discarded from further consideration. As evidenced by the data, single mutation variants MS-79 and MS-80 showed loss of activity for specific pseudoviruses. Also, the combinatorial variants MS-85 through MS-88 and the N-terminal variants MS-89 through MS-92 showed loss in activity and were removed from further consideration.
Example 4. Characterization of Round-2 PGDM1400 Variant Antibodies
[0328] Round-2 variants were designed based on the single light chain variants MS-66 (KV:F2I), MS-67 (KV:H9L), MS-69 (KV:S18P), MS-71 (KV:D73G), MS-73 (KV:T85A). The combinatorial variants built from these amino acid sets for the Round-2 variants are listed in Table 2. Assays used for analysis of the Round-2 variants included SEC to quantify monomer and HMW species following purification, DSF to characterize stability of the CH2 and Fab domains during thermal ramping, chemical unfolding, low pH stability, solubility, and retention of neutralization capacity.
[0329] Results of the initial screening consisting of SEC analysis for dimer and oligomer are shown in Table 11, while results for the DSF, low pH stability, chemical unfolding and solubility are shown in Tables 12 and 13. The dimer and oligomer content of all variants were similar to the parental molecule, MS-119 (Table 1), and were, thus, not a differentiating factor for identifying the optimal molecules. However, the DSF analysis demonstrated an increase of 3° C. for Tm2 of a number of variants, which also showed an increase in WSS by an average of 20 points. Conformational stability was evaluated by chemical unfolding, which assesses the intrinsic resistance of the native state against unfolding as measured by the mid-point of the denaturation curve. Variants with the highest Tm2 and WSS showed the greatest increase in inflection point (i.e., up to 0.25 M from the parental molecule) by chemical unfolding (Table 12). Interestingly, the presence of KV:F2I mutation was the common denominator across these variants. Variants containing the KV:F2I mutation showed an average WSS of 30.3 with the highest being 34 and the range being 26-34, compared to an average of 12.8 with a range of 11-15 for those combinatorial variants not containing the mutation. Additionally, the inflection point by chemical unfolding was higher in variants with the KV:F2I mutation (average value 2.44 M) compared to the parental molecule (average value 2.26 M), whereas, variants without the mutation showed an average value of 2.30 M. The fact that it takes slightly more GuHCl for the variant antibodies to reach the same point of chemical unfolding as the parental PGDM1400 antibody may be due to tighter packing of the hydrophobic core of the Fv. Together, these results are indicative of an increase in conformational stability of the combinatorial variants in comparison to the parental molecule.
[0330] Colloidal stability was also investigated and shown to increase for a number of the combinatorial variants (Table 13). Specifically, we investigated high temperature aggregation, solubility in PEG solutions and self-interaction nanoparticle spectroscopy (SINS). High temperature aggregation was investigated at 68° C. and 69.2° C., temperatures at which parental PGDM1400, MS-119, readily aggregates. Similar to the conformational stability results, a number of Round-2 combinatorial variants showed no aggregation at temperatures that cause aggregation of the parental PGDM1400 molecule. Again, the KV:F2I mutation was found to be central to these observations. Only those variants that carried the KV:F2I mutation were resistant to aggregation at high temperature, while variants without the mutation showed aggregation profiles similar to the parental molecule, MS-119. PEG solubility, which is indicative of protein/protein interaction was measured at 9.4% w/v PEG, a concentration at which only 50% of the parental molecule, MS-119 is soluble. The results for the solubility assay demonstrated a decrease in solubility for some variants, with a number of them being similar to the parental molecule, MS-119. This result was consistent with the method used to define destabilizing sites where solubility was not specifically targeted. Similarly, result from SINS analysis, which reports protein/protein interactions related to viscosity, was comparable between the variants and the parental molecule, MS-119, and are not shown.
[0331] Finally, the variants were assayed for retention of neutralization activity. Tables 14 and 15 show neutralization activity of Round-2 variants against 12 pseudoviruses of HIV that are representative of the broader set of viruses against which PGDM1400 is active. Antibodies with more than a 3-fold increase in the IC50 or IC80 value for a particular pseudovirus were considered inactive and discarded from further consideration. As evidenced from the data, the combinatorial variants showed similar IC50 and IC80 values within the approximate 3-fold limit of the assay.
[0332] Overall, analysis of the Round-2 variants (outlined in Tables 11-15) showed significant increase in multiple stability characteristics including thermal stability, chemical stability, and conformational stability, which are important for increased manufacturability and storage stability of the molecules.
TABLE-US-00006 TABLE 6 Analysis of biophysical characteristics of Round-1 PGDM1400 variant antibodies: titer and SEC Titer SEC SEC SEC Molecule Set (μg/ml) (% Main) (% Dimer) (% Oligomer) MS-119 56.5 88.62 8.77 2.61 MS-66 217 91.18 7.22 1.59 MS-67 124.2 91.52 7.18 1.3 MS-68 75.6 89.37 8.41 2.22 MS-69 169.8 91.25 7.34 1.41 MS-70 136.3 90.32 8.09 1.59 MS-71 182.2 91.8 7.03 1.17 MS-72 143.9 90.68 7.86 1.47 MS-73 213.1 90.89 7.71 1.4 MS-74 119.5 91.15 7.47 1.38 MS-75 113 90.07 8.17 1.76 MS-76 35 91.54 7.19 1.27 MS-77 106.7 91.09 7.36 1.55 MS-78 49 88.17 9.06 2.77 MS-79 39.7 89.56 8.54 1.9 MS-80 33.5 90.32 7.69 2 MS-81 39.3 88.7 8.75 2.55 MS-82 32.5 88.64 9 2.35 MS-83 92.8 87.42 10.17 2.42 MS-84 45.3 89.21 8.24 2.54 MS-85 187.3 90.51 7.8 1.69 MS-86 49.1 92.01 7 0.98 MS-87 44.6 90.73 7.72 1.55 MS-88 41.9 91.85 7.16 0.98 MS-89 77.6 92.12 6.81 1.07 MS-90 101.2 93.72 5.71 0.57 MS-91 89.5 93.57 5.64 0.79 MS-92 186.2 90.63 7.7 1.68
TABLE-US-00007 TABLE 7 Analysis of additional biophysical characteristics of Round-1 PGDM1400 variant antibodies: DSF and isothermal chemical unfolding DSF T1 DSF T2 Weighted Inflection Molecule ° C Std ° C. Std Shoulder Std Pt Std ΔG Std Set (Avg. n = 2) Dev (Avg. n = 2) Dev Score Dev (Avg n = 3) Dev (Avg n = 2) Dev MS-119 71.2 0.02 13.5 0.09 2.23 0.04 10.7 1.0 MS-66 70.4 0.21 77.5 0.07 36.12 4.13 2.38 0.02 14.5 3.2 MS-67 71.4 0.46 74.3 0.00 20.74 0.20 2.28 0.03 14.3 2.9 MS-68 70.4 0.13 11.91 0.02 2.02 0.03 8.2 0.6 MS-69 71.1 0.40 11.58 0.65 2.24 0.03 12.3 0.9 MS-70 71.4 0.22 73.2 0.00 15.17 0.60 2.28 0.03 12.7 1.3 MS-71 71.1 0.21 7.05 0.53 2.28 0.01 17.5 1.9 MS-72 71.0 0.13 12.66 0.00 2.19 0.07 10.7 2.4 MS-73 71.4 0.32 12.36 1.16 2.25 0.03 12.5 3.7 MS-74 71.5 0.36 12.26 0.21 2.23 0.02 11.2 2.3 MS-75 71.1 0.06 75.1 0.14 9.08 0.30 2.32 0.02 14.7 1.0 MS-76 71.1 0.22 17.43 3.51 2.21 0.02 12.4 3.0 MS-77 71.1 0.08 13.39 1.27 2.18 0.04 10.9 2.5 MS-78 71.1 0.19 15.07 0.14 2.34 0.02 14.3 2.6 MS-79 70.5 0.19 6.69 0.13 MS-80 69.7 0.03 7.59 0.40 2.02 0.02 7.3 1.5 MS-81 71.0 0.02 9.85 0.10 2.27 0.05 14.0 5.1 MS-82 70.5 0.02 10.96 0.57 MS-83 71.0 0.14 3.80 0.09 2.25 0.01 11.8 1.6 MS-84 69.8 0.12 13.74 0.39 MS-85 70.4 0.09 77.3 0.00 7.87 0.06 MS-86 71.0 0.17 75.2 0.00 13.34 0.54 2.32 0.04 13.7 3.6 MS-87 69.1 0.06 5.53 2.23 MS-88 69.6 0.06 32.36 1.24 2.12 0.03 9.4 1.5 MS-89 70.9 0.01 19.80 0.15 MS-90 71.0 0.17 75.4 0.00 23.47 0.63 2.32 0.01 14.1 2.1 MS-91 70.9 0.09 74.8 0.00 17.81 1.52 MS-92 70.5 0.04 78.4 0.14 29.87 0.57
TABLE-US-00008 TABLE 8 Analysis of additional biophysical characteristics of Round-1 PGDM1400 variant antibodies: low pH stability and PEG solubility pH 3.3 HMW % PEG Solubility Molecule Set (Avg n = 2) Std Dev (Avg n = 4) Std Dev MS-119 10.22 0.43 0.13 0.03 MS-66 5.62 0.26 0.11 0.03 MS-67 6.01 0.25 0.15 0.01 MS-68 6.74 0.08 0.14 0.02 MS-69 5.86 0.37 0.13 0.02 MS-70 5.46 0.22 0.15 0.01 MS-71 4.61 0.23 0.13 0.01 MS-72 6.35 0.54 0.12 0.01 MS-73 6.02 0.28 0.15 0.01 MS-74 5.84 0.21 0.14 0.01 MS-75 6.39 0.25 0.15 0.01 MS-76 6.27 0.14 0.11 0.01 MS-77 7.69 0.57 0.12 0.02 MS-78 7.94 0.47 0.13 0.01 MS-79 5.84 0.15 0.11 0.01 MS-80 3.56 0.13 0.10 0.01 MS-81 6.38 0.34 0.14 0.01 MS-82 6.36 0.26 0.10 0.01 MS-83 8.31 0.44 0.12 0.02 MS-84 8.18 0.47 0.11 0.01 MS-85 4.62 0.28 0.14 0.01 MS-86 4.63 0.82 0.10 0.01 MS-87 18.85 0.81 0.08 0.01 MS-88 5.06 0.06 0.06 0.01 MS-89 4.35 0.27 0.09 0.01 MS-90 4.80 0.17 0.09 0.02 MS-91 4.60 1.71 0.14 0.03 MS-92 5.62 0.26 0.12 0.01
TABLE-US-00009 TABLE 9 Analysis of neutralization activity of Round-1 PGDM1400 variant antibodies against representative PGDM1400 sensitive virus panel (SC422661.8, RHPA4259.7, Du172.17, BB1012-11.TC21, CNE52, 0260.v5.c36) in TZM.bl cells. Loss of potency are values > 3-fold of control value. Molecule SC422661.8 RHPA4259.7 Du172.17 BB1012-11.TC21 CNE52 0260.v5.c36 Set IC50 IC80 IC50 IC80 IC50 IC80 IC50 IC80 IC50 IC80 IC50 IC80 Control 0.872 4.865 0.340 1.197 2.380 9.384 0.031 0.102 0.452 2.727 0.035 0.138 MS-194 0.365 3.293 0.181 0.704 1.918 6.811 0.025 0.103 0.098 0.839 0.015 0.064 MS-66 0.902 5.820 0.238 0.745 2.076 11.315 0.017 0.074 0.408 3.743 0.024 0.092 MS-67 0.549 2.825 0.129 0.706 1.757 9.307 0.017 0.071 0.212 1.813 0.023 0.088 MS-68 0.538 2.021 0.199 0.759 1.821 6.797 0.023 0.076 0.136 1.133 0.019 0.073 MS-69 0.609 3.372 0.139 0.811 1.404 5.604 0.018 0.071 0.129 1.902 0.021 0.078 MS-70 0.391 2.430 0.182 0.734 1.493 6.053 0.021 0.079 0.301 2.547 0.028 0.103 MS-71 0.273 1.935 0.164 0.583 1.289 6.069 0.015 0.054 0.172 2.587 0.022 0.062 MS-72 0.373 2.826 0.247 1.157 1.748 8.758 0.023 0.089 0.302 4.534 0.023 0.114 MS-73 0.384 2.543 0.094 0.549 0.934 4.999 0.023 0.086 0.207 1.809 0.018 0.065 MS-74 0.388 3.506 0.132 0.461 1.460 5.570 0.021 0.093 0.169 1.457 0.017 0.086 MS-75 0.489 9.411 0.276 1.313 3.559 16.146 0.029 0.134 0.687 7.547 0.034 0.137 MS-76 0.360 6.457 0.242 0.842 1.702 9.675 0.013 0.059 0.174 1.372 0.010 0.060 MS-77 0.373 3.588 0.209 0.734 2.233 12.211 0.023 0.103 0.322 2.715 0.019 0.101 MS-78 0.635 3.631 0.241 0.841 1.714 6.997 0.024 0.105 0.233 1.394 0.020 0.095 MS-79 0.587 >25 0.221 0.792 7.279 >25 0.019 0.080 2.420 >25 0.028 0.159 MS-80 0.451 >25 0.267 1.240 15.238 >25 0.020 0.086 5.245 >25 0.041 0.321 MS-81 0.418 4.246 0.202 0.897 1.647 6.371 0.024 0.102 0.212 1.802 0.020 0.075 MS-82 0.387 3.295 0.192 0.672 1.881 6.995 0.023 0.108 0.206 1.183 0.016 0.051 MS-83 0.267 2.112 0.150 0.663 1.465 7.404 0.018 0.082 0.135 1.039 0.023 0.087 MS-84 0.192 1.374 0.175 0.773 1.635 6.554 0.019 0.085 0.092 0.830 0.016 0.062 MS-85 0.540 7.038 0.173 0.794 2.233 9.528 0.028 0.124 0.341 4.708 0.019 0.059 MS-86 1.196 >25 0.305 1.103 4.941 18.786 0.048 0.159 1.462 13.262 0.036 0.191 MS-87 0.711 13.927 0.294 1.093 >25 >25 0.023 0.102 2.686 >25 0.114 1.010 MS-88 1.410 >25 0.504 2.509 >25 >25 0.020 0.088 5.857 >25 0.178 1.514 MS-89 0.786 >25 0.290 1.040 11.866 >25 0.032 0.143 2.343 23.795 0.036 0.197 MS-90 1.480 >25 0.254 1.330 4.068 15.574 0.044 0.204 1.046 8.560 0.030 0.168 MS-91 1.655 22.127 0.267 1.824 3.851 23.865 0.037 0.170 1.405 10.577 0.046 0.171 MS-92 1.529 >25 0.250 1.155 6.850 >25 0.029 0.127 3.919 >25 0.044 0.350 Assay Set up: mAbs tested at primary concentration of 25 ug/ml and titrated 5-fold 7x (duplicate wells)
TABLE-US-00010 TABLE 10 Analysis of neutralization activity of Round-1 PGDM1400 variant antibodies against additional representative PGDM1400 sensitive virus panel (263-8, SC05.8C11.2344, X1193_c1, Ce1176_A3, AC10.0.29, 6952.v1.c20) in TZM.bl cells. Loss of potency are values > 3-fold of control value. Molecule 263-8 SC05.8C11.2344 X1193_c1 Ce1176_A3 AC10.0.29 6952.v1.c20 Set IC50 IC80 IC50 IC80 IC50 IC80 IC50 IC80 IC50 IC80 IC50 IC80 Control 0.018 0.085 0.575 2.533 0.148 0.525 0.252 2.788 0.051 0.230 0.600 10.550 MS-194 0.007 0.026 0.458 1.532 0.088 0.311 0.023 0.292 0.044 0.290 0.144 6.577 MS-66 0.017 0.060 0.811 2.536 0.086 0.322 0.165 5.427 0.035 0.213 0.284 >25 MS-67 0.011 0.037 0.631 2.144 0.075 0.303 0.105 1.898 0.031 0.186 0.280 7.888 MS-68 0.011 0.038 0.708 1.946 0.100 0.287 0.096 1.644 0.040 0.169 0.206 12.877 MS-69 0.010 0.046 0.456 1.690 0.072 0.326 0.087 1.709 0.029 0.169 0.182 3.064 MS-70 0.016 0.047 0.508 1.360 0.111 0.413 0.060 0.968 0.053 0.206 0.285 4.839 MS-71 0.014 0.041 0.425 1.410 0.072 0.265 0.040 0.796 0.035 0.177 0.325 7.924 MS-72 0.020 0.057 0.791 2.854 0.072 0.258 0.074 1.298 0.051 0.255 0.643 12.437 MS-73 0.010 0.032 0.461 1.624 0.085 0.327 0.035 0.581 0.041 0.206 0.222 6.993 MS-74 0.013 0.037 0.415 1.935 0.086 0.296 0.028 0.564 0.041 0.216 0.285 15.123 MS-75 0.018 0.086 0.792 3.718 0.174 0.591 0.139 2.785 0.055 0.442 0.977 >25 MS-76 0.015 0.053 0.587 2.567 0.126 0.575 0.082 1.631 0.047 0.368 0.844 >25 MS-77 0.012 0.033 0.516 2.412 0.109 0.398 0.067 1.120 0.036 0.311 0.236 6.741 MS-78 0.014 0.053 0.658 2.257 0.126 0.452 0.047 0.837 0.044 0.225 0.342 >25 MS-79 0.018 0.067 0.593 1.970 0.109 0.365 0.473 9.051 0.056 0.404 13.249 >25 MS-80 0.033 0.173 0.898 3.147 0.137 0.607 1.115 >25 0.054 0.280 >25 >25 MS-81 0.013 0.037 0.537 2.006 0.113 0.512 0.044 0.646 0.044 0.324 0.228 >25 MS-82 0.011 0.033 0.502 1.797 0.084 0.411 0.023 0.635 0.050 0.267 0.212 22.966 MS-83 0.007 0.050 0.381 1.879 0.061 0.308 0.029 0.405 0.038 0.259 0.126 3.089 MS-84 0.006 0.030 0.481 1.737 0.058 0.285 0.018 0.567 0.052 0.257 0.237 18.201 MS-85 0.011 0.043 0.425 1.488 0.068 0.507 0.073 1.719 0.044 0.242 0.393 >25 MS-86 0.009 0.050 1.622 5.393 0.167 0.371 0.366 4.246 0.107 0.741 2.548 >25 MS-87 0.065 0.361 1.231 5.167 0.098 0.277 0.907 24.985 0.059 0.328 >25 >25 MS-88 0.111 0.990 1.544 7.312 0.171 0.775 2.756 >25 0.089 0.743 >25 >25 MS-89 >25 >25 0.820 3.746 0.144 0.636 0.298 9.212 0.067 0.618 >25 >25 MS-90 0.021 0.109 1.496 3.841 0.114 0.399 0.220 4.240 0.059 0.592 2.678 >25 MS-91 0.020 0.075 0.848 2.838 0.057 0.388 0.213 2.042 0.070 0.632 2.615 >25 MS-92 0.024 0.208 1.537 4.092 0.110 0.529 0.600 9.783 0.051 0.446 7.376 >25
TABLE-US-00011 TABLE 11 Analysis of biophysical characteristics of Round-2 PGDM1400 variant antibodies: titer and SEC Titer SEC SEC SEC Molecule Set (μg/ml) (% Main) (% Dimer) (% Oligomer) MS-119 362.3 96.53 2.68 0.61 MS-93 529.4 96.46 2.81 0.55 MS-94 750.3 96.88 2.43 0.5 MS-95 831.4 85.9 12.98 0.86 MS-96 875.4 96.57 2.69 0.55 MS-97 160.9 95.71 3.29 0.69 MS-98 484.6 97.99 1.59 0.22 MS-99 322.2 96.7 2.64 0.44 MS-100 345.8 97.92 1.63 0.25 MS-101 411.2 97.01 2.36 0.43 MS-102 456.3 97.56 1.95 0.28 MS-103 430.7 97.12 2.23 0.46 MS-104 672.9 96.81 2.63 0.39 MS-105 631.5 95.68 3.28 0.84 MS-106 253.1 95.01 4.04 0.67 MS-107 312.7 94.22 4.63 0.93 MS-108 311.3 95.07 4.01 0.7 MS-109 278 96.66 2.68 0.45 MS-110 195.7 95.26 3.72 0.75 MS-111 444.2 97.1 2.33 0.4 MS-112 460.2 95.93 3.3 0.6 MS-113 343.6 94.62 4.41 0.75 MS-114 650 95.31 3.78 0.74 MS-115 541.9 95.28 3.87 0.67 MS-116 289.6 93.81 5.00 0.94 MS-117 509.9 96.02 3.24 0.58 MS-118 701.1 96.19 3.06 0.57
TABLE-US-00012 TABLE 12 Analysis of additional biophysical characteristics of Round-2 PGDM1400 variant antibodies: DSF, isothermal chemical unfolding and low pH stability Inflection Weighted Pt of pH 3.3 Molecule DSF T1° C. Std DSF T2° C. shoulder Std Unfolding Std HMW % Std Set (Avg. n = 2) Dev (Avg. n = 2) Std Dev Score Dev (Avg n = 3) Dev (Avg n = 2) Dev MS-119 70.6 0.1 74.2 0.1 13 0 2.26 0.02 3.74 0.13 MS-93 70.5 0.0 77.9 0.1 34 1 2.45 0.03 3.67 0.29 MS-94 70.3 0.0 77.4 0.1 29 1 2.39 0.02 3.97 0.83 MS-95 70.5 0.1 77.5 0.0 26 1 2.47 0.01 14.14 0.13 MS-96 70.5 0.1 77.7* 31 1 2.45 0.00 4.06 0.47 MS-97 70.7 0.3 74.0 0.1 14 1 2.28 0.04 4.90 0.42 MS-98 70.5 0.4 73.9 0.1 13 0 2.36 0.02 2.86 0.01 MS-99 70.3 0.0 74.5* 15 1 2.37 0.03 4.43 0.06 MS-100 70.5 0.2 73.5 0.1 11 1 2.27 0.03 3.22 0.04 MS-101 70.7 0.1 74.1 0.1 12 0 2.27 0.02 4.93 1.49 MS-102 70.6 0.1 73.9 0.2 12 0 2.33 0.01 3.90 1.58 MS-103 70.4 0.0 77.6 0.1 32 1 2.43 0.02 3.71 0.47 MS-104 70.6 0.0 77.8 0.0 31 0 2.50 0.01 3.28 0.42 MS-105 70.4 0.1 77.8 0.0 33 0 2.50 0.02 3.99 0.24 MS-106 70.4 0.0 77.3 0.0 27 1 2.40 0.01 4.65 0.21 MS-107 70.5 0.1 77.5 0.2 29 0 2.37 0.03 5.16 0.16 MS-108 70.5 0.0 77.5 0.1 29 1 2.43 0.01 4.83 0.06 MS-109 70.7 0.0 74.1 0.6 12 0 2.31 0.07 3.60 0.15 MS-110 70.7 0.1 73.8* 15 0 2.22 0.04 5.05 0.18 MS-111 70.9 0.4 74.0 0.1 14 0 2.36 0.03 2.73 0.15 MS-112 70.4 0.1 73.6 0.1 11 1 2.21 0.04 3.88 0.21 MS-113 70.5 0.1 77.6 0.0 31 0 2.46 0.02 4.53 0.18 MS-114 70.4 0.1 77.7 0.1 32 0 2.40 0.01 3.84 0.12 MS-115 70.5 0.1 77.8 0.0 32 0 2.52 0.02 3.93 0.14 MS-116 70.6 0.1 77.4 0.4 28 1 2.40 0.02 5.25 0.13 MS-117 70.8 0.0 74.0 0.1 12 0 2.32 0.03 3.22 0.11 MS-118 70.6 0.1 77.6 0.1 30 1 2.46 0.03 3.33 0.13 *Only one of two DSF analysis showed a Tm2 value
TABLE-US-00013 TABLE 13 Analysis of additional biophysical characteristics of Round-2 PGDM1400 variant antibodies: thermal hold, solubility, and SINS Thermal Hold: Thermal Hold: 9.4% PEG Molecule A350 Heated A350 Heated Solubility Std Set 68° C. in 69.2° C. in (Avg. n = 4) Dev MS-119 0.5211 0.5941 0.14 0.01 MS-93 0.0798 0.0953 0.13 0.02 MS-94 0.0798 0.2821 0.13 0.01 MS-95 0.0807 0.1843 0.12 0.01 MS-96 0.0756 0.1181 0.14 0.01 MS-97 0.6173 0.4481 0.13 0.01 MS-98 0.5878 0.5949 0.13 0.01 MS-99 0.5764 0.6219 0.14 0.01 MS-100 0.6164 0.64 0.11 0.01 MS-101 0.6221 0.5229 0.13 0.01 MS-102 0.5824 0.6174 0.12 0.01 MS-103 0.0694 0.1403 0.14 0.02 MS-104 0.0703 0.1336 0.13 0.01 MS-105 0.0904 0.5936 0.15 0.01 MS-106 0.0946 0.392 0.12 0.01 MS-107 0.0996 0.2462 0.13 0.02 MS-108 0.0916 0.2408 0.12 0.01 MS-109 0.6081 0.5371 0.12 0.01 MS-110 0.623 0.504 0.13 0.01 MS-111 0.5932 0.5551 0.13 0.01 MS-112 0.6375 0.5974 0.13 0.01 MS-113 0.0885 0.3335 0.12 0.01 MS-114 0.0882 0.1156 0.14 0.02 MS-115 0.0808 0.1305 0.13 0.01 MS-116 0.0863 0.3428 0.12 0.01 MS-117 0.6592 0.5923 0.11 0.02 MS-118 0.0812 0.1959 0.13 0.01
TABLE-US-00014 TABLE 14 Analysis of neutralization activity of selected Round-2 PGDM1400 variant antibodies against representative PGDM1400 sensitive virus panel (SC422661.8, RHPA4259.7, Du172.17, BB1012-11.TC21, CNE52, 0260.v5.c36) in TZM.bl cells. Loss of potency are values > 3-fold of control value. Molecule SC422661.8 RHPA4259.7 DU172.17 BB1012-11.TC21 CNE52 0260.v5.c36 Set IC50 IC80 IC50 IC80 IC50 IC80 IC50 IC80 IC50 IC80 IC50 IC80 Control 0.398 1.954 0.406 1.097 3.234 9.188 0.057 0.157 0.609 5.122 0.040 0.143 MS-194 0.194 0.975 0.369 1.693 2.144 5.980 0.032 0.089 0.259 2.183 0.027 0.095 MS-93 0.333 2.705 0.358 1.605 3.240 9.142 0.034 0.118 0.636 5.599 0.034 0.116 MS-94 0.315 1.229 0.358 0.985 3.027 11.204 0.055 0.146 0.455 5.180 0.027 0.095 MS-95 0.433 2.883 0.208 0.690 1.970 7.068 0.044 0.117 0.567 5.048 0.022 0.060 MS-96 0.397 2.817 0.252 1.137 3.579 12.966 0.047 0.195 0.543 5.358 0.026 0.093 MS-97 0.311 2.089 0.183 0.824 2.213 7.535 0.040 0.130 0.214 1.622 0.028 0.096 MS-98 0.301 2.136 0.146 0.676 1.441 6.335 0.025 0.114 0.237 1.907 0.019 0.068 MS-99 0.294 1.456 0.257 0.718 1.949 7.141 0.032 0.084 0.219 1.798 0.024 0.087 MS-100 0.184 1.309 0.142 0.694 1.562 5.255 0.029 0.095 0.147 2.015 0.019 0.053 MS-101 0.251 1.876 0.227 0.816 2.410 6.878 0.037 0.121 0.145 1.231 0.025 0.069 MS-102 0.170 1.207 0.178 0.649 1.161 5.910 0.031 0.100 0.211 1.869 0.019 0.053 MS-103 0.408 1.986 0.290 1.328 2.681 10.194 0.046 0.155 0.432 4.179 0.030 0.104 MS-104 0.277 1.520 0.227 1.021 2.578 9.413 0.032 0.137 0.509 6.483 0.024 0.084 MS-105 0.440 3.509 0.270 1.245 3.058 11.527 0.038 0.124 0.441 5.037 0.032 0.110 MS-106 0.201 1.075 0.143 0.720 1.487 8.035 0.031 0.132 0.420 4.307 0.018 0.066 MS-107 0.362 2.070 0.219 0.949 2.576 13.931 0.035 0.111 0.389 5.290 0.034 0.109 MS-108 0.148 1.250 0.178 1.103 1.714 13.591 0.027 0.110 0.265 3.505 0.028 0.088 MS-109 0.233 1.749 0.182 0.794 1.144 8.759 0.021 0.087 0.206 1.951 0.027 0.089 MS-110 0.243 1.191 0.180 0.817 0.847 7.298 0.033 0.104 0.210 1.308 0.026 0.088 MS-111 0.285 1.028 0.178 0.802 1.761 6.270 0.029 0.093 0.220 1.306 0.024 0.066 MS-112 0.275 1.435 0.195 0.661 1.772 4.908 0.032 0.129 0.133 1.155 0.022 0.059 MS-113 0.354 2.116 0.206 0.696 2.223 8.271 0.026 0.109 0.502 5.045 0.026 0.092 MS-114 0.236 2.210 0.237 1.244 1.658 7.738 0.033 0.136 0.444 3.607 0.030 0.105 MS-115 0.202 1.030 0.237 0.647 1.833 5.035 0.024 0.106 0.489 5.459 0.025 0.067 MS-116 0.185 1.063 0.187 0.663 1.973 6.710 0.030 0.139 0.321 3.090 0.024 0.067 MS-117 0.168 1.332 0.201 0.715 1.908 6.489 0.030 0.099 0.206 1.726 0.021 0.057 MS-118 0.244 2.053 0.164 0.785 2.117 8.294 0.036 0.117 0.420 3.400 0.019 0.069 Assay Set up: mAbs tested at primary concentration of 25 ug/ml and titrated 5-fold 7x (duplicate wells).
TABLE-US-00015 TABLE 15 Analysis of neutralization activity of selected Round-2 PGDM1400 variant antibodies against additional representative PGDM1400 sensitive virus panel (263-8, SC05.8C11.2344, X1193_c1, Ce1176_A3, AC10.0.29, 6952.v1.c20) in TZM.bl cells. Loss of potency are values > 3-fold of control value. Molecule 263-8 SC05.8C11.2344 X1193_c1 Ce1176_A3 AC10.0.29 6952.v1.c20 Set IC50 IC80 IC50 IC80 IC50 IC80 IC50 IC80 IC50 IC80 IC50 IC80 Control 0.019 0.066 0.781 2.641 0.121 0.779 0.517 13.569 0.073 0.368 0.257 3.988 MS-194 0.012 0.045 0.710 2.254 0.072 0.456 0.189 3.835 0.057 0.219 1.560 3.014 MS-93 0.018 0.065 0.749 2.483 0.088 0.543 0.239 5.326 0.058 0.301 0.259 4.307 MS-94 0.014 0.047 0.739 2.505 0.123 0.548 0.371 11.686 0.079 0.299 0.238 3.670 MS-95 0.013 0.047 0.702 1.789 0.111 0.467 0.203 5.384 0.076 0.270 0.229 6.261 MS-96 0.018 0.064 1.072 3.397 0.113 0.677 0.248 5.848 0.064 0.253 0.558 8.561 MS-97 0.015 0.052 1.022 2.630 0.101 0.588 0.125 1.594 0.057 0.282 0.147 0.917 MS-98 0.009 0.033 0.590 1.897 0.088 0.369 0.102 2.361 0.067 0.245 0.109 1.596 MS-99 0.011 0.041 0.759 2.492 0.130 0.570 0.074 1.252 0.072 0.328 0.217 1.993 MS-100 0.006 0.026 0.531 1.769 0.058 0.358 0.048 0.905 0.042 0.211 0.182 1.676 MS-101 0.011 0.041 0.819 2.686 0.098 0.602 0.043 1.942 0.045 0.221 0.254 3.509 MS-102 0.007 0.027 0.527 1.418 0.071 0.412 0.039 2.747 0.047 0.168 0.162 2.527 MS-103 0.018 0.070 0.707 2.436 0.112 0.502 0.172 3.134 0.070 0.334 0.485 5.952 MS-104 0.014 0.052 0.512 1.726 0.063 0.388 0.218 6.109 0.041 0.208 0.457 8.608 MS-105 0.017 0.062 0.794 2.725 0.067 0.417 0.172 3.393 0.051 0.394 0.706 9.489 MS-106 0.011 0.039 0.411 1.439 0.093 0.394 0.099 3.432 0.062 0.299 0.216 3.799 MS-107 0.016 0.055 0.658 2.281 0.130 0.570 0.147 3.986 0.079 0.397 0.457 8.867 MS-108 0.012 0.042 0.428 1.450 0.083 0.545 0.117 3.843 0.045 0.216 0.323 3.366 MS-109 0.010 0.033 0.546 1.852 0.073 0.467 0.078 5.834 0.040 0.199 0.188 1.744 MS-110 0.010 0.038 0.914 3.100 0.106 0.677 0.097 2.496 0.076 0.285 0.195 2.120 MS-111 0.016 0.045 0.496 1.694 0.109 0.352 0.082 3.121 0.061 0.258 0.213 2.097 MS-112 0.013 0.037 0.501 1.675 0.069 0.293 0.068 2.033 0.056 0.262 0.160 1.467 MS-113 0.018 0.052 0.573 1.942 0.098 0.421 0.182 6.054 0.052 0.245 0.351 5.030 MS-114 0.018 0.066 0.810 3.860 0.124 0.404 0.139 7.391 0.066 0.313 0.146 1.369 MS-115 0.013 0.037 0.436 1.514 0.080 0.274 0.180 6.301 0.049 0.249 0.303 4.700 MS-116 0.011 0.042 0.456 1.617 0.064 0.222 0.145 3.116 0.052 0.258 0.341 7.508 MS-117 0.010 0.029 0.432 1.482 0.064 0.226 0.113 2.173 0.051 0.252 0.208 3.840 MS-118 0.011 0.039 0.449 1.538 0.085 0.287 0.260 8.123 0.056 0.273 0.392 12.192 Assay Set up: mAbs tested at primary concentration of 25 μg/ml and titrated 5-fold 7x (duplicate wells)
Example 5. Pharmacokinetic Characterization of PGDM1400 Variant Antibodies
[0333] Mice were injected with PGDM1400 variant antibodies and pharmacokinetic properties of the variants was tested in blood samples collected up to 28 days (e.g., up to about 1 hour, 2 hour, 3 hour, 4 hour, 5 hour, 6 hour, 7 hour, 8 hour, 9 hour, 10 hour, 11 hour, 12 hour, 13 hour, 14 hour 15 hour, 16 hour, 17 hour, 18 hour, 19 hour, 20 hour, 21 hour, 22 hour, 23 hour, 1 day, 2 day, 3 day, 4 day, 5 day, 6 day, 7 day, 8 day, 9 day, 10 day, 11 day, 12 day, 13 day, 14 day, 15 day, 16 day, 17 day, 18 day, 19 day, 20 day, 21 day, 22 day, 23 day, 24 day, 25 day, 26 day, 27 day, or 28 day) post-infusion. Infusion and sample collection were done as per the schedule outlined in Table 16.
TABLE-US-00016 TABLE 16 Dosing and blood sampling schedule 3BNC117 Test mAb TA Dose, Route, # of Mouse Group (Pettit 650) mg/kg Frequency mice Strain Blood Sampling Time 1 MS-65: PGDM1400 10 IV, 1X 4 Tg276 1 h, 8 h, 2 d, 5 d, 7d, 10 d, 14 d, 21 d, 28 d 2 MS-119: PGDM1400-LS 10 IV, 1X 4 Tg276 1 h, 8 h, 2 d, 5 d, 7 d, 10 d, 14 d, 21 d, 28 d 3 MS-93: Optimized 10 IV, 1X 4 Tg27S6 1 h, 8 h, 2 d, 5 d, 7 d, 10 d, PGDM1400-LS 14 d, 21 d, 28 d 4 MS-103: Optimized 10 IV, 1X 4 Tg276 1 h, 8 h, 2 d, 5 d, 7 d, 10 d, PGDM1400-LS 14 d, 21 d, 28 d 5 MS-115: Optimized 10 IV, 1X 4 Tg276 1 h, 8 h, 2 d, 5 d, 7 d, 10 d, PGDM1400-LS 14 d, 21 d, 28 d
[0334] Pharmacokinetics of PGDM1400 variant antibodies was studied by antibody binding assays that were adapted from validated BAMA (binding assay multiplex assay) for detection of antibodies specific for HIV-1 antigens. The assays were done in 96-well plates using beads coupled to neutravidin and bound to biotinylated mouse anti-human IgG Fc antibody. Infused monoclonal antibody (mAb) was detected with an antibody to the human Ig Kappa chain. Blood samples (up to 28 day post-infusion) were tested at 1:200, 1:500, 1:1000, and 1:2000 dilutions. All samples, standards and controls were tested in duplicate and several samples were tested in 2 separate assays to confirm observed concentration. Samples were received in plates arranged by mAb variant received and timepoint post infusion, and diluted at 1:10. Standard curves for each mAb were titrated in assay diluent and applied in a 5PL (five parameter logistic) curve algorithm to determine the concentration of the corresponding infused mAb variant. Standard curve EC50's were tracked in Levey Jennings charts against historical means obtained from development assays. Controls included blank wells, blank (no antigen) beads, and antigen-specific controls.
[0335] Results from the binding assays demonstrated that the parental PGDM1400 anti-ID antibody did not have equal affinity for the PGDM1400 variant antibodies (
TABLE-US-00017 TABLE 17 Concentration of antibody in post-infusion blood sample Average Concentration of Responders (μg/ml) MS-93: MS-103: MS-115: Hours Post MS-65: MS-119: Optimized Optimized Optimized Infusion PGDM1400 PGDM1400-LS PGDM1400-LS PGDM1400-LS PGDM1400-LS 1 164.39 148.10 161.72 199.86 144.68 8 90.72 109.28 108.16 133.67 98.09 24 48.45 76.75 74.86 94.88 73.40 48 32.49 66.06 60.70 73.70 58.50 120 11.83 54.07 43.82 53.96 49.54 168 6.13 46.41 42.03 45.52 30.91 240 1.00 30.00 28.15 29.75 4.57 336 0.47 23.89 24.47 20.09 504 13.75 15.31 9.40 672 8.50 9.44 3.69
Example 6. Treatment of a Subject with a PGDM1400 Variant Antibody or Antigen-Binding Fragment Thereof
[0336] One or more PGDM1400 variant antibodies or antigen-binding fragments thereof described herein, or a composition containing the same can be administered to a subject, such as a human (e.g., a HIV-infected human or a human at risk of HIV transmission) in order to treat or prevent HIV infection (e.g., HIV-1 infection). Administration of the one or more PGDM1400 variant antibodies or antigen-binding fragments thereof or a composition containing the same, for instance, can reduce proviral DNA (e.g., to below about 1,000 DNA copies/10.sup.6 cells or to an undetectable level) in a tissue (e.g., lymph node tissue, gastrointestinal tissue, and/or peripheral blood), decrease plasma viral load (e.g., to less than 3,500 RNA copies/ml or to an undetectable level), increase HIV-specific cell-mediated immune response and/or humoral immune response, and/or decrease viral replication in the subject. For instance, an HIV-infected human can be treated by administering one or more PGDM1400 variant antibodies or antigen-binding fragments thereof described herein or a composition containing the same by an appropriate route (e.g., intravenously) at a particular dosage (e.g., about 0.01-5000 mg or about 0.01-100 mg/kg of the antibody or antigen-binding fragment thereof) one or more times daily, weekly, every two weeks, every three weeks, or monthly. A single dose or more than one dose of the one or more PGDM1400 variant antibodies or antigen-binding fragments thereof described herein or a composition containing the same can be administered to the subject over a course of days, weeks, months, or years.
[0337] The progression of HIV infection that is treated with the PGDM1400 variant antibody or antigen-binding fragment thereof described herein or a composition containing the same can be monitored by any one or more of several established methods. A physician can monitor the subject by direct observation in order to evaluate how the symptoms exhibited by the subject have changed in response to treatment (e.g., by evaluation of proviral DNA, plasma viral load and/or viral replication in the subject). Based on such observations, a physician may prescribe higher/lower dosages or more/less frequent dosing of the PGDM1400 variant antibody or antigen-binding fragment or a composition containing the same in subsequent rounds of treatment.
Example 7. Treatment of a Subject with a PGDM1400 Variant Antibody or Antigen-Binding Fragment Thereof in Combination with an Immunotherapy Agent
[0338] The PGDM1400 variant antibody or antigen-binding fragment described herein or a composition containing the same (e.g., MS-93, MS-94, MS-95, MS-96, MS-103, MS-104, MS-105, MS-106, MS-107, MS-108, MS-113, MS-114, MS-115, MS-116, and MS-118) can be administered to a subject, such as a human (e.g., a HIV-infected human or a human at risk of HIV transmission) in combination with (for instance, admixed with, co-administered with, or administered separately from) one or more: (i) immunomodulators (e.g., AS-101, Bropirimine, Acemannan, CL246,738, EL10, FP-21399, Gamma Interferon, Granulocyte Macrophage Colony Stimulating Factor, HIV Core Particle Immunostimulant, IL-2, Immune Globulin Intravenous, IMREG-1, IMREG-2, Imuthiol Diethyl Dithio Carbamate, Alpha-2 Interferon, Methionine-Enkephalin, MTP-PE Muramyl-Tripeptide, Granulocyte Colony Stimulating Factor, Remune, CD4 (e.g., recombinant soluble CD4), rCD4-IgG hybrids, SK&F106528 Soluble T4, Thymopentin, Tumor Necrosis Factor, or Infliximab); (ii) reservoir activators, such as a PKC agonist (e.g., a phorbol ester, a macrocyclic lactone such as bryostatin-1, or a diterpene such as an ingenol compound), a cytokine or chemokine (e.g., interleukin (IL)-7, IL-15, or interferon-alpha (IFN-α)), a Toll-like receptor (TLR) agonist (e.g., a TLR 1/2 agonist (e.g., Pam3CSK4), a TLR3 agonist (e.g., Poly-ICLC), a TLR5 agonist (e.g., flagellin), a TLR7 agonist (e.g., GS-9620), or a TLR9 agonist (e.g., MGN1703 and CpG7909)), an immune checkpoint inhibitor (e.g., anti-PD-1 monoclonal antibody, an anti-PD-1 ligand (PD-L1) monoclonal antibody, or an anti-CTLA-4 monoclonal antibody), a histone deacetylase (HDAC) inhibitor (e.g., romidepsin, vorinostat, belinostat, LAQ824, panobinostat, entinostat, C1994, or mocetinostat), or a small molecule reservoir activator (e.g., disulfiram, a benzotriazole derivative (e.g., 3-Hydroxy-1,2,3-benzotriazin-4((3H)-one (HO-DHBt); a SMAC mimetic), or a BRG-Brahma Associated Factor (BAF) inhibitor (e.g., caffeic acid phenethyl ester or pyrimethamine)); (iii) antiretroviral agent (ARV) (e.g., lamivudine and zidovudine, emtricitabine (FTC), zidovudine (ZDV), azidothymidine (AZT), lamivudine (3TC), zalcitabine, dideoxycytidine (ddC), tenofovir disoproxil fumarate (TDF), didanosine (ddl), stavudine (d4T), abacavir sulfate (ABC), etravirine, delavirdine (DLV), efavirenz (EFV), nevirapine (NVP), amprenavir (APV), tipranavir (TPV), indinavir (IDV), saquinavir, saquinavir mesylate (SQV), lopinavir (LPV), ritonavir (RTV), fosamprenavir calcium (FOS-APV), ritonavir, RTV, darunavir, atazanavir sulfate (ATV), nelfinavir mesylate (NFV), enfuvirtide, T-20, maraviroc, raltegravir, ibalizumab, IL-2, IL-12, or alpha-epibromide); and/or one, two, three, or more different HIV-specific broadly neutralizing antibodies (bnAb), such as a CD4 binding site (CD4bs)-specific antibody (e.g., 3BNC117 or VRC07-523), an N332 glycan-dependent antibody (e.g., PGT121), or a V2-specific antibody (e.g., CAP256-VRC26 or PGDM1400). The one or more immunomodulator(s), reservoir activator(s), ARV(s), and/or HIV-specific bnAb(s) can be administered prior to (e.g., 1 year, 9 months, 6 months, 3 months, 1 month, 3 weeks, 2 weeks, 1 week, 5 days, 3 days, 1 day, 18 hours, 12 hours, 6 hours, or 1 hour prior to), concurrently with and/or after (e.g., 1 year, 9 months, 6 months, 3 months, 1 month, 3 weeks, 2 weeks, 1 week, 5 days, 3 days, 1 day, 18 hours, 12 hours, 6 hours, or 1 hour after) the administration of the PGDM1400 variant antibody or antigen-binding fragment described herein or a composition containing the same. Administration routes, dosage and frequency of administration of the PGDM1400 variant antibody or antigen-binding fragment or a composition containing the same has been exemplified in the aforementioned Example 6.
[0339] The progression of HIV infection that is treated with the PGDM1400 variant antibody or antigen-binding fragment thereof in combination with the one or more immunomodulator(s), reservoir activator(s), ARV(s), and/or HIV-specific bnAb(s) can be monitored by any one or more of several established methods. A physician can monitor the subject by direct observation in order to evaluate how the symptoms exhibited by the subject have changed in response to treatment (e.g., by evaluation of proviral DNA, plasma viral load and/or viral replication in the subject). Based on such observations, a physician may prescribe higher/lower dosages or more/less frequent dosing of the PGDM1400 variant antibody or antigen-binding fragment or a composition containing the same in combination with the one or more immunomodulator(s), reservoir activator(s), ARV(s), and/or HIV-specific bnAb(s) in subsequent rounds of treatment.
Other Embodiments
[0340] While the invention has been described in connection with specific embodiments thereof, it will be understood that it is capable of further modifications and this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure come within known or customary practice within the art to which the invention pertains and may be applied to the essential features hereinbefore set forth.
[0341] All publications, patents, and patent applications are herein incorporated by reference in their entirety to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated by reference in its entirety.