PROCESSES FOR SEPARATING AGGREGATED PROTEINS FROM MONOMERIC PROTEINS IN A BIOLOGICAL SOLUTION

Abstract

A process for separating aggregated proteins from monomeric proteins in a biological solution, the process including: providing at least one filter element having a contacting surface, wherein the filter element comprises filter media comprising: a porous substrate; and disposed on the porous substrate, a polymer comprising a hydrocarbon backbone and a plurality of pendant groups attached to the hydrocarbon backbone, wherein each of a first plurality of pendant groups comprises: (1) at least one acidic group or salt thereof; and (2) a spacer group that directly links the at least one acidic group or salt thereof to the hydrocarbon backbone by a chain of at least 6 catenated atoms; and allowing an initial biological solution to contact the contacting surface of the filter element N under conditions effective to separate the aggregated proteins from the monomeric proteins such that a final biological solution includes purified monomeric proteins.

Claims

1. A process for separating aggregated proteins from monomeric proteins in a biological solution, the process comprising: providing at least one filter element having a contacting surface, wherein the filter element comprises filter media comprising: a porous substrate; and disposed on the porous substrate, a polymer comprising a hydrocarbon backbone and a plurality of pendant groups attached to the hydrocarbon backbone, wherein each of a first plurality of pendant groups comprises: (1) at least one acidic group or salt thereof; and (2) a spacer group that directly links the at least one acidic group or salt thereof to the hydrocarbon backbone by a chain of at least 6 catenated atoms; and allowing an initial biological solution to contact the contacting surface of the filter element under conditions effective to separate the aggregated proteins from the monomeric proteins such that a final biological solution includes purified monomeric proteins.

2. The process of claim 1 wherein the polymer comprises interpolymerized units of at least one monomer comprising: (1) at least one ethylenically unsaturated group; (2) at least one acidic group or salt thereof; and (3) a spacer group that directly links the at least one ethylenically unsaturated group and the at least one acidic group or salt thereof by a chain of at least 6 catenated atoms.

3. The process of claim 1 further comprising recovering the monomeric proteins without eluting.

4. The process of claim 1 wherein the conditions are effective to recover, in the final solution, at least 70% of the monomeric proteins present in the initial biological solution.

5. The process of claim 1 wherein the conditions are effective to remove at least 10% of the aggregated proteins from the initial biological solution.

6. The process of claim 1 wherein the proteins comprise antibodies, enzymes, or hormones.

7. The process of claim 6 wherein the proteins comprise antibodies.

8. The process of claim 1 wherein the conditions comprise a pH of the biological solution of below 9.

9. The process of claim 1 wherein the conditions comprise a conductivity of the biological solution of at least 1 mS/cm.

10. The process of claim 1 wherein the at least one acidic group or salt thereof of the polymer disposed on the porous substrate is present at a density of at least 0.02 mmole/gram of filter media.

11. The process of claim 2 wherein the at least one ethylenically unsaturated group is selected from an ethenyl group, a 1-alkylethenyl group, and a combination thereof.

12. The process of claim 1 wherein the chain of the spacer group has at least 8 catenated atoms.

13. The process of claim 1 wherein the at least one acidic group or salt thereof is selected from a carboxy group, a phosphono group, a phosphato group, a sulfono group, a sulfato group, a boronato group, and a combination thereof.

14. The process of claim 1 wherein the spacer group is a catenated heteroatom-containing hydrocarbon group.

15. The process of claim 1 wherein the spacer group comprises at least one hydrogen bonding moiety.

Description

EXAMPLES

[0250] Objects and advantages of this disclosure are further illustrated by the following examples, but the particular materials and amounts thereof recited in these examples, as well as other conditions and details, should not be construed to unduly limit this disclosure. These examples are merely for illustrative purposes only and are not meant to be limiting on the scope of the appended claims.

Materials:

[0251]

TABLE-US-00001 TABLE 1 Materials Description (abbreviation) Source 2-[Bis(2-hydroxyethyl)amino]-2- Sigma-Aldrich, (hydroxymethyl)propane-1,3-diol (Bis-Tris) St. Louis, MO 2-(N-morpholino)ethanesolfonic acid (MES) Alfa Aesar, Ward Hill, MA Sodium phosphate (monobasic and dibasic) JT Baker, Phillipsburg, NJ Tris(hydroxymethyl)-aminomethane (Tris) Alfa Aesar, Ward Hill, MA Sodium citrate Alfa Aesar, Ward Hill, MA Citric acid Alfa Aesar, Ward Hill, MA Vinyldimethylazlactone (VDM) SNPE, Inc., Princeton, NJ 2-Isocyanatoethyl methacrylate (IEM) Showa Denko KK, Kanagawa, Japan 4-(3-Sulfopropyloxy)benzophenone, Described in Japanese sodium salt (S-BP) Patent No. 47040913 (Teijin Ltd.)

IgG Antibody Solution Preparation:

[0252] Monoclonal antibody IgG (33.2 mg/mL, 3-4% aggregate content, pI>8, pH 5.3, 3.0 mS/cm) was diluted either 10-fold or 40-fold with a 20 mM buffer solution of citrate (pH 5 and 6), MES (pH 6), Bis-Tris (pH 6), sodium phosphate (pH 7), or Tris (pH 8). The resulting buffer solutions were adjusted with sodium chloride to have conductivities ranging between 1.5-105 mS/cm. The conductivity and pH measurements of the IgG-containing buffer solutions were determined using an Accumet Excel XL50 conductivity meter (Fisher Scientific, Hampton, N.H.) and a VWR sympHony benchtop pH meter (VWR, Radnor, Pa.), respectively. In Table 2, the buffer, pH and conductivity for each of the IgG-containing buffer solutions (IgG Solution Numbers 1-27) are recorded.

TABLE-US-00002 TABLE 2 IgG-containing solutions used for examples IgG Solution Buffer Conductivity Number (20 mM) pH (mS/cm) 1 Citrate 5 3.4 2 Citrate 5 10.5 3 Citrate 5 22.1 4 Citrate 5 35.0 5 Citrate 5 99.7 6 Citrate 6 2.8 7 Citrate 6 9.9 8 Citrate 6 20.5 9 Citrate 6 36.8 10 Citrate 6 98.3 11 MES 6 1.7 12 MES 6 9.0 13 MES 6 20.1 14 MES 6 35.9 15 Bis-Tris 6 2.5 16 Bis-Tris 6 11.9 17 Bis-Tris 6 22.5 18 Phosphate 7 3.0 19 Phosphate 7 10.2 20 Phosphate 7 20.4 21 Phosphate 7 36.2 22 Phosphate 7 101.4 23 Tris 8 2.1 24 Tris 8 9.6 25 Tris 8 21.1 26 Tris 8 35.5 27 Tris 8 97.5

Monomer Preparation:

Monomer Example A.

4-[[[2-Methyl-2-(prop-2-enoylamino)propanoyl]amino]methyl]cyclohexanecarboxylic acid, sodium salt (VDM-4-aminomethyl-cyclohexanecarboxylic acid, sodium salt)

[0253] ##STR00036##

[0254] Trans-4-Aminomethyl-cyclohexanecarboxylic acid (2.00 g, 0.013 mol) was added to a 100 mL round bottom flask. An aqueous solution of sodium hydroxide (1.0 N, 13 mL) was added to the flask and the resulting mixture was stirred until the solids dissolved. The flask was then placed in an ice-water bath and stirred for 15 minutes. 2-Vinyl-4,4-dimethylazlactone (VDM) (1.77 g, 1.7 mL, 0.013 mol) was added by syringe and the reaction was stirred for 30 minutes with the flask continuously maintained in the ice-water bath. The cooling bath was then removed and the reaction was allowed to warm to room temperature over a period of 30 minutes. The pH of the reaction was adjusted to about 7 by the addition of a few drops of a concentrated hydrochloric acid solution. .sup.1H-NMR of an aliquot confirmed the formation of 4-[[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]methyl]cyclohexanecarboxylic acid, sodium salt. .sup.1H-NMR (D.sub.2O, 500 MHz) 6 0.89 (q, 2H), 1.27 (q, 2H), 1.42 (s, 6H), 1.43 (m, 1H), 1.69 (d, 2H), 1.82 (d, 2H), 2.03 (m, 1H), 2.98 (d, 2H), 5.72 (m, 1H), 6.14 (m, 1H), 6.23 (m, 1H).

Monomer Example B.

2-Hydroxy-4-2-methyl-2-(prop-2-enoylamino)propanoylaminobutanoic acid, sodium salt (VDM-2-hydroxy-4-aminobutanoic acid, sodium salt)

[0255] ##STR00037##

[0256] 2-Hydroxy-4-aminobutanoic acid (5.00 g, 0.042 mol) was added to a 100 mL round bottom flask. An aqueous solution of sodium hydroxide solution (1.0 N, 42 mL) was added to the flask and the resulting mixture was stirred until the solids dissolved. The flask was then placed in an ice-water bath and stirred for 15 minutes. VDM (5.84 g, 5.6 mL, 0.042 mol) was added by syringe and the reaction was stirred for 30 minutes with the flask continuously maintained in the ice-water bath. The cooling bath was then removed and the reaction was allowed to warm to room temperature over a period of 30 minutes. .sup.1H-NMR of an aliquot confirmed the formation of 2-hydroxy-4-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]butanoic acid, sodium salt. The pH of the reaction was adjusted to about 7 by the addition of a few drops of a concentrated hydrochloric acid solution. .sup.1H-NMR (D.sub.2O, 500 MHz) 1.43 (s, 6H), 1.71 (m, 1H), 1.89 (m, 1H), 3.22 (m, 1H), 3.29 (m, 1H), 3.99 (m, 2H), 5.73 (m, 1H), 6.15 (m, 1H), 6.23 (m, 1H).

Monomer Example C.

3-Hydroxy-2-2-methyl-2-(prop-2-enoylamino)propanoylaminopropanoic acid, sodium salt (VDM-serine, sodium salt)

[0257] ##STR00038##

[0258] L-Serine (5.00 g, 0.048 mol) was added to a 100 mL round bottom flask. An aqueous solution of sodium hydroxide (1.0 N, 48 mL) was added to the flask and the resulting mixture was stirred until the solids dissolved. The flask was then placed in an ice-water bath and stirred for 15 minutes. VDM (6.62 g, 6.4 mL, 0.048 mol) was added by syringe and the reaction was stirred for 30 minutes with the flask continuously maintained in the ice-water bath. The cooling bath was then removed and the reaction was allowed to warm to room temperature over a period of 30 minutes. The pH of the reaction was adjusted to about 7 by the addition of a few drops of a concentrated hydrochloric acid solution. .sup.1H-NMR of an aliquot confirmed the formation of 3-hydroxy-2-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]propanoic acid, sodium salt. .sup.1H-NMR (D.sub.2O, 500 MHz) 1.48 (s, 6H), 3.79 (m, 2H), 4.22 (m, 1H), 5.74 (m, 1H), 6.16 (m, 1H), 6.25 (m, 1H).

Monomer Example D.

3-(4-Hydroxyphenyl)-2-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]propanoic acid, sodium salt (VDM-tyrosine, sodium salt)

[0259] ##STR00039##

[0260] DL-Tyrosine (5.00 g, 0.028 mol) was added to a 250 mL round bottom flask. An aqueous solution of sodium hydroxide (0.5 N, 150 mL) was added to the flask and the resulting mixture was stirred until the solids dissolved. The flask was then placed in an ice-water bath and stirred for 15 minutes. VDM (3.84 g, 3.7 mL, 0.028 mol) was added by syringe and the reaction was stirred for 2 hours with the flask continuously maintained in the ice-water bath. The cooling bath was then removed and the reaction was allowed to warm to room temperature over a period of 30 minutes. .sup.1H-NMR of an aliquot confirmed the formation of 3-(4-hydroxyphenyl)-2-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]propanoic acid, sodium salt. .sup.1H-NMR (D.sub.2O, 500 MHz) 1.39 (s, 6H), 2.70 (m, 1H), 2.84 (m, 1H), 3.42 (m, 1H), 5.67 (m, 1H), 6.11 (m, 1H), 6.22 (m, 1H), 6.58 (m, 2H), 6.95 (m, 2H).

Monomer Example E.

3-(1H-Indol-3-yl)-2-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]propanoic acid, sodium salt (VDM-tryptophan, sodium salt)

[0261] ##STR00040##

[0262] DL-Tryptophan (5.00 g, 0.025 mol) was added to a 250 mL round bottom flask. An aqueous solution of sodium hydroxide solution (0.5 N, 100 mL) was added to the flask and the resulting mixture was stirred until the solids dissolved. The flask was then placed in an ice-water bath and stirred for 15 minutes. VDM (3.40 g, 3.5 mL, 0.025 mol) was added by syringe and the reaction was stirred for 3 hours with the flask continuously maintained in the ice-water bath. The cooling bath was then removed and the reaction was allowed to warm to room temperature over a period of 30 minutes. .sup.1H-NMR of an aliquot confirmed the formation of 3-(1H-indol-3-yl)-2-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]propanoic acid, sodium salt. .sup.1H-NMR (D.sub.2O, 500 MHz) 1.39 (s, 6H), 3.01 (m, 1H), 3.13 (m, 1H), 3.54 (m, 1H), 5.67 (m, 1H), 6.11 (m, 1H), 6.21 (m, 1H), 7.12 (m, 1H), 7.19 (m, 2H), 7.46 (m, 1H), 7.70 (m, 1H).

Monomer Example F.

7-[[2-Methyl-2-(prop-2-enoylamino)propanoyl]amino]heptanoic acid, sodium salt (VDM-7-aminoheptanoic acid, sodium salt)

[0263] ##STR00041##

[0264] 7-Aminoheptanoic acid (5.00 g, 0.035 mol) was added to a 100 mL round bottom flask. An aqueous solution of sodium hydroxide (1.0 N, 35 mL) was added to the flask and the resulting mixture was stirred until the solids dissolved. The flask was then placed in an ice-water bath and stirred for 15 minutes. VDM (4.79 g, 4.6 mL, 0.035 mol) was added by syringe and the reaction was stirred for 30 minutes with the flask continuously maintained in the ice-water bath. The cooling bath was then removed and the reaction was allowed to warm to room temperature over a period of 30 minutes. The pH of the reaction was adjusted to about 7 by the addition of a few drops of a concentrated hydrochloric acid solution. .sup.1H-NMR of an aliquot confirmed the formation of 7-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]heptanoic acid, sodium salt. .sup.1H-NMR (D.sub.2O, 500 MHz) 1.24 (s, 6H), 1.41 (m, 8H), 1.49 (m, 2H), 2.11 (m, 2H), 3.11 (m, 2H), 5.72 (m, 1H), 6.15 (m, 1H), 6.22 (m, 1H).

Monomer Example G.

3-(1H-Imidazol-5-yl)-2-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]propanoic acid, sodium salt (VDM-histidine, sodium salt)

[0265] ##STR00042##

[0266] L-Histidine (2.00 g, 0.013 mol) was added to a 100 mL round bottom flask. An aqueous solution of sodium hydroxide 1.0 N, 13 mL) was added to the flask and the resulting mixture was stirred until the solids dissolved. The flask was then placed in an ice-water bath and stirred for 15 minutes. VDM (1.79 g, 1.72 mL, 0.013 mol) was added by syringe and the reaction was stirred for 30 minutes with the flask continuously maintained in the ice-water bath. The cooling bath was then removed and the reaction was allowed to warm to room temperature over a period of 30 minutes. .sup.1H-NMR of an aliquot confirmed the formation of 3-(1H-imidazol-5-yl)-2-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]propanoic acid, sodium salt. The pH of the reaction was adjusted to about 7 by the addition of a few drops of a concentrated hydrochloric acid solution. .sup.1H-NMR (D.sub.2O, 500 MHz) 1.30 (d, 6H), 2.87 (dd, 1H), 3.04 (dd, 1H), 4.24 (q, 1H), 5.61 (d, 1H), 6.01 (d, 1H), 6.12 (dd, 1H), 6.90 (s, 1H), 7.92 (s, 1H).

Monomer Example H.

3-[2-(2-Methylprop-2-enoyloxy)ethylcarbamoylamino]propanoic acid, sodium salt (IEM-3-aminopropanoic acid, sodium salt)

[0267] ##STR00043##

[0268] 3-Aminopropanoic acid (1.78 g, 0.02 mol) was added to a 100 mL round bottom flask. An aqueous solution of sodium hydroxide solution (1.0 N, 20 mL) was added to the flask and the resulting mixture was stirred until the solids dissolved. The flask was then placed in an ice-water bath and stirred for 15 minutes. 2-Isocyanatoethyl methacrylate (IEM) (3.1 g, 0.02 mol) was added by syringe and the reaction was stirred for 30 minutes with the flask continuously maintained in the ice-water bath. The cooling bath was then removed and the reaction was allowed to warm to room temperature over a period of 30 minutes. A colorless precipitate was filtered from the reaction mixture. The pH of the filtrate was adjusted to about 7 by the addition of a few drops of a concentrated hydrochloric acid solution. .sup.1H-NMR of an aliquot of the filtrate confirmed the formation of 3-[2-(2-methylprop-2-enoyloxy)ethylcarbamoylamino]propanoic acid, sodium salt. .sup.1H-NMR (D.sub.2O, 500 MHz) 1.78 (s, 3H), 2.22 (t, 2H), 3.16 (t, 2H), 3.30 (t, 2H), 4.07 (t, 2H), 5.58 (s, 1H), 5.99 (s, 1H).

Monomer Example I.

2-[2-(2-Methylprop-2-enoyloxy)ethylcarbamoylamino]ethanesulfonic acid, sodium salt (IEM-taurine, sodium salt)

[0269] ##STR00044##

[0270] Taurine (2.50 g, 0.02 mol) was added to a 100 mL round bottom flask. An aqueous solution of sodium hydroxide solution (1.0 N, 20 mL) was added to the flask and the resulting mixture was stirred until the solids dissolved. The flask was then placed in an ice-water bath and stirred for 15 minutes. IEM (3.1 g, 0.02 mol) was added by syringe and the reaction was stirred for 30 minutes with the flask continuously maintained in the ice-water bath. The cooling bath was then removed and the reaction was allowed to warm to room temperature over a period of 30 minutes. A colorless precipitate was filtered from the reaction mixture. The pH of the filtrate was adjusted to about 7 by the addition of a few drops of a concentrated hydrochloric acid solution. .sup.1H-NMR of an aliquot of the filtrate confirmed the formation of 2-[2-(2-methylprop-2-enoyloxy)ethylcarbamoylamino]ethanesulfonic acid, sodium salt. .sup.1H-NMR (D.sub.2O, 500 MHz) 1.75 (s, 3H), 2.88 (t, 2H), 3.28 (t, 2H) 3.32 (t, 2H), 4.06 (t, 2H), 5.56 (m, 1H), 5.97 (s, 1H).

Monomer Example J.

2-[[2-Methyl-2-(prop-2-enoylamino)propanoyl]amino]ethanesulfonic acid, sodium salt (VDM-taurine, sodium salt)

[0271] ##STR00045##

[0272] Taurine (2.50 g, 0.02 mol) was added to a 100 mL round bottom flask. An aqueous solution of sodium hydroxide solution (1.0 N, 20 mL) was added to the flask and the resulting mixture was stirred until the solids dissolved. The flask was then placed in an ice-water bath and stirred for 15 minutes. VDM (2.78 g, 0.02 mol) was added by syringe and the reaction was stirred for 30 minutes with the flask continuously maintained in the ice-water bath. The cooling bath was then removed and the reaction was allowed to warm to room temperature over a period of 30 minutes. The pH of the reaction was adjusted to about 7 by the addition of a few drops of a concentrated hydrochloric acid solution. .sup.1H-NMR of an aliquot confirmed the formation of 2-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]ethanesulfonic acid, sodium salt. .sup.1H-NMR (D.sub.2O, 500 MHz) 1.35 (s, 6H), 2.94 (t, 2H), 3.45 (t, 2H) 5.64 (m, 1H), 6.10 (m, 2H).

Monomer Example K.

2-[2-Methyl-2-(prop-2-enoylamino)propanoyl]oxyethylphosphonic acid (VDM-2-(hydroxyethyl)phosphonic acid)

[0273] ##STR00046##

[0274] 2-Hydroxyethyl dimethylphosphate (90% purity, 15.0 g, 97.3 mmol) and 1,8-diazabicyclo5.4.0undec-7-ene (1 mL) were added to a 250 mL flask containing a solution of VDM (13.7 g, 97.3 mmol) dissolved in 100 mL of anhydrous methylene chloride and the reaction was stirred overnight. The reaction mixture was diluted with additional methylene chloride and washed sequentially with saturated 5% NaH.sub.2PO.sub.4, H.sub.2O, and brine. The organic phase was dried with Na.sub.2SO.sub.4 and then filtered. A small amount (approximately 130 mg) of BHT was added and the solution was concentrated under reduced pressure to give 11.2 g of a colorless liquid. The liquid was dissolved in 50 mL of methylene chloride and placed in ice-water bath under a nitrogen atmosphere. Bromotrimethylsilane (12.3 g, 80.2 mmol) was added and the reaction was stirred for 90 minutes at room temperature. The solution was concentrated under reduced pressure. Methanol (50 mL) was added to the resulting residue and the solution was stirred at room temperature for 1 hour. The solution was then concentrated under reduced pressure at room temperature to yield the final product as clear liquid. .sup.1H-NMR of an aliquot confirmed the formation of 2-[2-methyl-2-(prop-2-enoylamino)propanoyl]oxyethylphosphonic acid. NMR (D.sub.2O, 500 MHz) 1.44 (s, 6H), 2.14 (dt, 2H), 4.29 (dt, 2H), 5.69 (dd, 1H), 6.12 (dd, 1H), 6.19 (broad s, 1H).

Monomer Example L.

3-[[2-Methyl-2-(prop-2-enoylamino)propanoyl]amino]propanoic acid, sodium salt (VDM-3-aminopropanoic acid, sodium salt)

[0275] ##STR00047##

[0276] 3-Aminopropanoic acid (1.78 g, 0.02 mol) was added to a 100 mL round bottom flask. An aqueous solution of sodium hydroxide (1.0 N, 20 mL) was added to the flask and the resulting mixture was stirred until the solids dissolved. The flask was then placed in an ice-water bath and stirred for 15 minutes. VDM (2.78 g, 0.02 mol) was added by syringe and the reaction was stirred for 30 minutes with the flask continuously maintained in the ice-water bath. The cooling bath was then removed and the reaction was allowed to warm to room temperature over a period of 30 minutes. The pH of the reaction was adjusted to about 7 by the addition of a few drops of a concentrated hydrochloric acid solution. .sup.1H-NMR of an aliquot confirmed the formation of 3-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]propanoic acid, sodium salt. .sup.1H-NMR (D.sub.2O, 500 MHz) 1.32 (s, .sup.6H), 2.23 (t, 2H), 3.24 (t, 2H), 5.63 (m, 1H), 6.0-6.2 (m, 2H).

Monomer Example M.

4-[[2-Methyl-2-(prop-2-enoylamino)propanoyl]amino]butanoic acid, sodium salt (VDM-GABA, sodium salt)

[0277] ##STR00048##

[0278] 4-Aminobutanoic acid (2.06 g, 0.02 mol) was added to a 100 mL round bottom flask. An aqueous solution of sodium hydroxide (1.0 N, 20 mL) was added to the flask and the resulting mixture was stirred until the solids dissolved. The flask was then placed in an ice-water bath and stirred for 15 minutes. VDM (2.78 g, 0.02 mol) was added by syringe and the reaction was stirred for 30 minutes with the flask continuously maintained in the ice-water bath. The cooling bath was then removed and the reaction was allowed to warm to room temperature over a period of 30 minutes. The pH of the reaction was adjusted to about 7 by the addition of a few drops of a concentrated hydrochloric acid solution. .sup.1H-NMR of an aliquot confirmed the formation of 4-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]butanoic acid, sodium salt. .sup.1H-NMR (D.sub.2O, 500 MHz) 1.34 (s, 6H), 1.59 (p, 2H), 2.04 (t, 2H), 3.05 (t, 2H), 5.62 (d, 1H), 6.0-6.2 (m, 2H).

Monomer Example N.

5-[[2-Methyl-2-(prop-2-enoylamino)propanoyl]amino]pentanoic acid, sodium salt (VDM-5-aminovaleric acid, sodium salt)

[0279] ##STR00049##

[0280] 5-Aminopentanoic acid (2.34 g, 0.02 mol) was added to a 100 mL round bottom flask. An aqueous solution of sodium hydroxide (1.0 N, 20 mL) was added to the flask and the resulting mixture was stirred until the solids dissolved. The flask was then placed in an ice-water bath and stirred for 15 minutes. VDM (2.78 g, 0.02 mol) was added by syringe and the reaction was stirred for 30 minutes with the flask continuously maintained in the ice-water bath. The cooling bath was then removed and the reaction was allowed to warm to room temperature over a period of 30 minutes. The pH of the reaction was adjusted to about 7 by the addition of a few drops of a concentrated hydrochloric acid solution. .sup.1H-NMR of an aliquot confirmed the formation of 5-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]pentanoic acid, sodium salt. .sup.1H-NMR (D.sub.2O, 500 MHz) 1.33 (s and m, 10H), 2.04 (t, 2H), 3.05 (t, 2H), 5.63 (d, 1H), 6.0-6.2 (m, 2H).

Monomer Example O.

6-[[2-Methyl-2-(prop-2-enoylamino)propanoyl]amino]hexanoic acid, sodium salt (VDM-6-aminocaproic acid, sodium salt)

[0281] ##STR00050##

[0282] 6-Aminohexanoic acid (2.62 g, 0.02 mol) was added to a 100 mL round bottom flask. An aqueous solution of sodium hydroxide (1.0 N, 20 mL) was added to the flask and the resulting mixture was stirred until the solids dissolved. The flask was then placed in an ice-water bath and stirred for 15 minutes. VDM (2.78 g, 0.02 mol) was added by syringe and the reaction was stirred for 30 minutes with the flask continuously maintained in the ice-water bath. The cooling bath was then removed and the reaction was allowed to warm to room temperature over a period of 30 minutes. The pH of the reaction was adjusted to about 7 by the addition of a few drops of a concentrated hydrochloric acid solution. .sup.1H-NMR of an aliquot confirmed the formation of 6-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]hexanoic acid, sodium salt. .sup.1H-NMR (D.sub.2O, 500 MHz) 1.14 (m, 2H), 1.34 (s and m, 8H), 1.41 (m, 2H), 2.02 (t, 2H), 3.03 (t, 2H), 5.62 (d, 1H), 6.0-6.2 (m, 2H).

Monomer Example P.

[0283] 2-[[2-Methyl-2-(prop-2-enoylamino)propanoyl]amino]-3-phenylpropanoic acid, sodium salt (VDM-phenylalanine, sodium salt)

##STR00051##

[0284] L-Phenylalanine (3.3 g, 0.02 mol) was added to a 100 mL round bottom flask. An aqueous solution of sodium hydroxide (1.0 N, 20 mL) was added to the flask and the resulting mixture was stirred until the solids dissolved. The flask was then placed in an ice-water bath and stirred for 15 minutes. VDM (2.78 g, 0.02 mol) was added by syringe and the reaction was stirred for 30 minutes with the flask continuously maintained in the ice-water bath. The cooling bath was then removed and the reaction was allowed to warm to room temperature over a period of 30 minutes. The pH of the reaction was adjusted to about 7 by the addition of a few drops of a concentrated hydrochloric acid solution. .sup.1H-NMR of an aliquot confirmed the formation of 2-2-methyl-2-(prop-2-enoylamino)propanoylamino-3-phenylpropanoic acid, sodium salt. .sup.1H-NMR (D.sub.2O, 500 MHz) 1.26 (s, 6H), 2.89 (m, 1H), 2.95 (m, 1H), 4.30 (m, 1H), 5.62 (d, 1H), 6.00-6.10 (m, 2H), 7.07-7.20 (m, 5H).

Monomer Example Q.

2-[2-(2-Methylprop-2-enoyloxy)ethylcarbamoylamino]-3-phenylpropanoic acid, sodium salt (IEM-phenylalanine, sodium salt)

[0285] ##STR00052##

[0286] L-Phenylalanine (3.3 g, 0.02 mol) was charged to a 100 mL round bottom flask. An aqueous solution of sodium hydroxide (1.0 N, 20 mL) was added to the flask and the resulting mixture was stirred until the solids dissolved. The flask was then placed in an ice-water bath and stirred for 15 minutes. IEM (3.1 g, 0.02 mol) was added by syringe and the reaction was stirred for 30 minutes with the flask continuously maintained in the ice-water bath. The cooling bath was then removed and the reaction was allowed to warm to room temperature over a period of 30 minutes. A colorless precipitate was filtered from the reaction mixture. The pH of the filtrate was adjusted to about 7 by the addition of a few drops of a concentrated hydrochloric acid solution. .sup.1H-NMR of an aliquot of the filtrate confirmed the formation of 2-[2-(2-methylprop-2-enoyloxy)ethylcarbamoylamino]-3-phenylpropanoic acid, sodium salt. .sup.1H-NMR (D.sub.2O, 500 MHz) 1.74 (br. s, 3H), 2.73 (m, 1H), 2.99 (m, 1H), 3.13 (m, 1H), 3.26 (m 1H), 3.90 (m, 2H), 4.17 (m, 1H), 5.54 (m, 1H), 5.95 (m, 1H), 7.09 and 7.15 (m, 5H).

Monomer Example R.

2-[2-(2-Methylprop-2-enoyloxy)ethylcarbamoylamino]ethanoic acid, sodium salt (IEM-glycine, sodium salt)

[0287] ##STR00053##

[0288] Glycine (1.5 g, 0.02 mol) was charged to a 100 mL round bottom flask. An aqueous solution of sodium hydroxide (1.0 N, 20 mL) was added to the flask and the resulting mixture was stirred until the solids dissolved. The flask was then placed in an ice-water bath and stirred for 15 minutes. IEM (3.1 g, 0.02 mol) was added by syringe and the reaction was stirred for 30 minutes with the flask continuously maintained in the ice-water bath. The cooling bath was then removed and the reaction was allowed to warm to room temperature over a period of 30 minutes. A colorless precipitate was filtered from the reaction mixture. The pH of the filtrate was adjusted to about 7 by the addition of a few drops of a concentrated hydrochloric acid solution. .sup.1H-NMR of an aliquot of the filtrate confirmed the formation of 2-[2-(2-methylprop-2-enoyloxy)ethylcarbamoylamino]ethanoic acid, sodium salt. .sup.1H-NMR (D.sub.2O, 500 MHz) 1.79 (s, 3H), 3.33 (m, 2H), 3.54 (s, 2H), 4.09 (m, 2 H), 5.59 (s, 1H), 5.99 (s, 1H).

Example 1

[0289] A coating solution was prepared by mixing a solution of 4-[[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]methyl]cyclohexanecarboxylic acid, sodium salt (Monomer Example A) (3.34 grams of a 23.85% w/w solution in deionized water, 2.5 mmol) with the initiator 4-(3-sulfopropyloxy)benzophenone, sodium salt (S-BP) (500 L of a 0.05 g/mL solution in deionized water). This mixture was diluted to a total of 5 grams with deionized water to produce a final monomer concentration of 0.5M and initiator concentration of 0.5%. A nylon membrane substrate (9 cm12 cm; nylon 66 membrane, single reinforced layer nylon three-zone membrane, nominal pore size 1.8 m, #080ZN, obtained from 3M Purification, Inc., Meridan, Conn.) was placed on a sheet of clear polyester film (about 0.25 mm thick), and the coating solution was pipetted onto the top surface of the substrate. The coating solution was allowed to soak into the substrate for about 1 minute, and then a second sheet of clear polyester film (about 0.25 mm thick) was placed over the top surface of the substrate. A 2.28 kg cylindrical weight was rolled over the top of the resulting three-layer sandwich (polyester film-membrane substrate-polyester film) to squeeze out excess coating solution. Ultraviolet (UV)-initiated grafting was conducted by irradiating the sandwich using a UV stand (Classic Manufacturing, Inc., Oakdale, Minn.) equipped with 18 bulbs (Sylvania RG2 40W F40/350BL/ECO, 10 above and 8 below the substrate, 1.17 meters (46 inches) long, spaced 5.1 cm (2 inches) on center), with an irradiation time of 15 minutes. The polyester sheets were removed, and the remaining functionalized substrate was placed in a 250 mL polyethylene bottle. The bottle was filled with 0.9 percent (wt %) saline, sealed, and shaken for 30 minutes to wash off any residual monomer or ungrafted polymer. The saline was decanted, and the functionalized substrate was washed for another 30 minutes with fresh saline solution and then washed for 30 minutes with deionized water and allowed to dry. The finished filter element had a graft density of 0.20 mmol/gram. Graft density was determined by the measuring the mass gain after conversion of the substrate to the finished filter element.

Example 2

[0290] A coating solution was prepared by mixing a solution of 2-hydroxy-4-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]butanoic acid, sodium salt (Monomer Example B) (2.98 grams of a 23.55% w/w solution in deionized water, 2.5 mmol) with S-BP (500 L of a 0.05 g/mL solution in deionized water). This mixture was diluted to a total of 5 grams with deionized water to produce a final monomer concentration of 0.5M and initiator concentration of 0.5%. The procedure described in Example 1 was followed to provide a finished filter element with a graft density of 0.07 mmol/gram.

Example 3

[0291] A coating solution was prepared by mixing a solution of 3-hydroxy-2-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]propanoic acid, sodium salt (Monomer Example C) (2.79 grams of a 23.85% w/w solution in deionized water, 2.5 mmol) with S-BP (500 L of a 0.05 g/mL solution in deionized water). This mixture was diluted to a total of 5 grams with deionized water to produce a final monomer concentration of 0.5M and initiator concentration of 0.5%. The procedure described in Example 1 was followed to provide a finished filter element with a graft density of 0.23 mmol/gram.

Example 4

[0292] A coating solution was prepared by mixing a solution of 3-(1H-imidazol-5-yl)-2-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]propanoic acid, sodium salt (Monomer Example G) (2.87 grams of a 27.55% w/w solution in deionized water, 2.5 mmol) with S-BP (500 L of a 0.05 g/mL solution in deionized water). This mixture was diluted to a total of 5 grams with deionized water to produce a final monomer concentration of 0.5M and initiator concentration of 0.5%. The procedure described in Example 1 was followed to provide a finished filter element with a graft density of 0.19 mmol/gram.

Example 5

[0293] A coating solution was prepared by mixing a solution of 4-[[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]methyl]cyclohexanecarboxylic acid, sodium salt (Monomer Example A) (1.67 grams of a 23.85% w/w solution in deionized water, 1.25 mmol) with S-BP (500 L of a 0.05 g/mL solution in deionized water). This mixture was diluted to a total of 5 grams with deionized water to produce a final monomer concentration of 0.25M and initiator concentration of 0.5%. The procedure described in Example 1 was followed to provide a finished filter element with a graft density of 0.12 mmol/gram.

Example 6

[0294] A coating solution was prepared by mixing a solution of 2-hydroxy-4-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]butanoic acid, sodium salt (Monomer Example B) (1.48 grams of a 23.55% w/w solution in deionized water, 1.25 mmol) with (S-BP (500 L of a 0.05 g/mL solution in deionized water). This mixture was diluted to a total of 5 grams with deionized water to produce a final monomer concentration of 0.25M and initiator concentration of 0.5%. The procedure described in Example 1 was followed to provide a finished filter element with a graft density of 0.04 mmol/gram.

Example 7

[0295] A coating solution was prepared by mixing a solution of 3-hydroxy-2-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]propanoic acid, sodium salt (Monomer Example C) (1.39 grams of a 23.85% w/w solution in deionized water, 1.25 mmol) with S-BP (500 L of a 0.05 g/mL solution in deionized water). This mixture was diluted to a total of 5 grams with deionized water to produce a final monomer concentration of 0.25M and initiator concentration of 0.5%. The procedure described in Example 1 was followed to provide a finished filter element with a graft density of 0.11 mmol/gram.

Example 8

[0296] A coating solution was prepared by mixing a solution of 3-(1H-imidazol-5-yl)-2-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]propanoic acid, sodium salt (Monomer Example G) (1.45 grams of a 27.55% w/w solution in deionized water, 1.25 mmol) with S-BP (500 L of a 0.05 g/mL solution in deionized water). This mixture was diluted to a total of 5 grams with deionized water to produce a final monomer concentration of 0.25M and initiator concentration of 0.5%. The procedure described in Example 1 was followed to provide a finished filter element with a graft density of 0.08 mmol/gram.

Example 9

[0297] A coating solution was prepared by mixing a solution of 2-hydroxy-4-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]butanoic acid, sodium salt (Monomer Example B) (2.98 grams of a 23.55% w/w solution in deionized water, 2.5 mmol) with S-BP (100 L of a 0.05 g/mL solution in deionized water). This mixture was diluted to a total of 5 grams with deionized water to produce a final monomer concentration of 0.5M and initiator concentration of 0.1%. The procedure described in Example 1 was followed to provide a finished filter element with a graft density of 0.10 mmol/gram.

Example 10

[0298] A coating solution was prepared by mixing a solution of 4-[[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]methyl]cyclohexanecarboxylic acid, sodium salt (Monomer Example A) (1.67 grams of a 23.85% w/w solution in deionized water, 1.25 mmol) with S-BP (100 L of a 0.05 g/mL solution in deionized water). This mixture was diluted to a total of 5 grams with deionized water to produce a final monomer concentration of 0.25M and initiator concentration of 0.1%. The procedure described in Example 1 was followed to provide a finished filter element with a graft density of 0.07 mmol/gram.

Example 11

[0299] A coating solution was prepared by mixing a solution of 3-hydroxy-2-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]propanoic acid, sodium salt (Monomer Example C) (1.39 grams of a 23.85% w/w solution in deionized water, 1.25 mmol) with S-BP (100 L of a 0.05 g/mL solution in deionized water). This mixture was diluted to a total of 5 grams with deionized water to produce a final monomer concentration of 0.25M and initiator concentration of 0.1%. The procedure described in Example 1 was followed to provide a finished filter element with a graft density of 0.07 mmol/gram.

Example 12

[0300] A coating solution was prepared by mixing a solution of 3-(4-hydroxyphenyl)-2-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]propanoic acid, sodium salt (Monomer Example D) (1.25 mL of a 1M solution in deionized water) with N,N-Dimethylacrylamide (1.25 mL of a 1M solution), and S-BP (500 L of a 0.05 g/mL solution in deionized water). This mixture was diluted to a total of 5 grams with deionized water to produce a final monomer concentration of 0.25M and initiator concentration of 0.5%. The procedure described in Example 1 was followed to provide a finished filter element with a graft density of 0.21 mmol/gram.

Example 13

[0301] A coating solution was prepared by mixing a solution of 3-(1H-indol-3-yl)-2-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]propanoic acid, sodium salt (monomer Example E) (1.25 mL of a 1M solution in deionized water) with S-BP (500 L of a 0.05 g/mL solution in deionized water). This mixture was diluted to a total of 5 grams with deionized water to produce a final monomer concentration of 0.25M and initiator concentration of 0.5%. The procedure described in Example 1 was followed to provide a finished filter element with a graft density of 0.16 mmol/gram.

Example 14

[0302] A coating solution was prepared by mixing a solution of 2-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]ethanesulfonic acid, sodium salt (Monomer Example J) (2.5 mL of a 1M solution in deionized water) with S-BP (500 L of a 0.05 g/mL solution in deionized water). This mixture was diluted to a total of 5 grams with deionized water to produce a final monomer concentration of 0.5M and initiator concentration of 0.5%. The procedure described in Example 1 was followed to provide a finished filter element with a graft density of 0.15 mmol/gram.

Example 15

[0303] A coating solution was prepared by mixing a solution of 2-2-methyl-2-(prop-2-enoylamino)propanoyloxyethylphosphonic acid, sodium salt (Monomer Example K) (2.5 mL of a 1M solution in deionized water) with S-BP (500 L of a 0.05 g/mL solution in deionized water). This mixture was diluted to a total of 5 grams with deionized water to produce a final monomer concentration of 0.5M and initiator concentration of 0.5%. The procedure described in Example 1 was followed to provide a finished filter element with a graft density of 0.20 mmol/gram.

Example 16

[0304] A coating solution was prepared by mixing a solution of 3-[2-(2-methylprop-2-enoyloxy)ethylcarbamoylamino]propanoic acid, sodium salt (monomer Example H) (3.75 mL of a 1M solution in deionized water) with S-BP (500 L of a 0.05 g/mL solution in deionized water). This mixture was diluted to a total of 5 grams with deionized water to produce a final monomer concentration of 0.75M and initiator concentration of 0.5%. The procedure described in Example 1 was followed to provide a finished filter element with a graft density of 0.50 mmol/gram.

Example 17

[0305] A coating solution was prepared by mixing a solution of 3-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]propanoic acid, sodium salt (Monomer Example L) (3.75 mL of a 1M solution in deionized water) with S-BP (500 L of a 0.05 g/mL solution in deionized water). This mixture was diluted to a total of 5 grams with deionized water to produce a final monomer concentration of 0.75M and initiator concentration of 0.5%. The procedure described in Example 1 was followed to provide a finished filter element with a graft density of 0.623 mmol/gram.

Example 18

[0306] A coating solution was prepared by mixing a solution of 4-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]butanoic acid, sodium salt (Monomer Example M) (1.53 grams of a 21.65% w/w solution in deionized water, 1.25 mmol) with S-BP (125 L of a 0.05 g/mL solution in deionized water). This mixture was diluted to a total of 5 grams with deionized water to produce a final monomer concentration of 0.25M and initiator concentration of 0.125%. The procedure described in Example 1 was followed to provide a finished filter element with a graft density of 0.23 mmol/gram.

Example 19

[0307] A coating solution was prepared by mixing a solution of 5-[[2-methyl-2-(prop-2-enoylamino)propanoyl]aminolpentanoic acid, sodium salt (Monomer Example N) (1.58 grams of a 22.05% w/w solution in deionized water, 1.25 mmol) with S-BP (62.5 L of a 0.05 g/mL solution in deionized water). This mixture was diluted to a total of 5 grams with deionized water to produce a final monomer concentration of 0.25M and initiator concentration of 0.0625%. The procedure described in Example 1 was followed to provide a finished filter element with a graft density of 0.19 mmol/gram.

Example 20

[0308] A coating solution was prepared by mixing a solution of 6-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]hexanoic acid, sodium salt (Monomer Example O) (1.54 grams of a 23.7% w/w solution in deionized water, 1.25 mmol) with S-BP (31.2 L of a 0.05 g/mL solution in deionized water). This mixture was diluted to a total of 5 grams with deionized water to produce a final monomer concentration of 0.25M and initiator concentration of 0.0312%. The procedure described in Example 1 was followed to provide a finished filter element with a graft density of 0.20 mmol/gram.

Example 21

[0309] A coating solution was prepared by mixing a solution of 7-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]heptanoic acid, sodium salt (Monomer Example F) (1.56 grams of a 24.5% w/w solution in deionized water, 1.25 mmol) with S-BP (62.5 L of a 0.05 g/mL solution in deionized water). This mixture was diluted to a total of 5 grams with deionized water to produce a final monomer concentration of 0.25M and initiator concentration of 0.0625%. The procedure described in Example 1 was followed to provide a finished filter element with a graft density of 0.18 mmol/gram.

Example 22

[0310] A coating solution was prepared by mixing a solution of 7-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]heptanoic acid, sodium salt (Monomer Example F) (1.56 grams of a 24.5% w/w solution in deionized water, 1.25 mmol) with S-BP (125 L of a 0.05 g/mL solution in deionized water). This mixture was diluted to a total of 5 grams with deionized water to produce a final monomer concentration of 0.25M and initiator concentration of 0.125%. The procedure described in Example 1 was followed to provide a finished filter element with a graft density of 0.24 mmol/gram.

Example 23

[0311] A coating solution was prepared by mixing a solution of 2-2-methyl-2-(prop-2-enoylamino)propanoyl]aminol-3-phenylpropanoic acid, sodium salt (monomer Example P) (1.65 grams of a 24.8% w/w solution in deionized water, 1.25 mmol) with S-BP (31.2 L of a 0.05 g/mL solution in deionized water). This mixture was diluted to a total of 5 grams with deionized water to produce a final monomer concentration of 0.25M and initiator concentration of 0.0312%. The procedure described in Example 1 was followed to provide a finished filter element with a graft density of 0.18 mmol/gram.

Example 24

[0312] A coating solution was prepared by mixing a solution of 2-2-(2-methylprop-2-enoyloxy)ethylcarbamoylamino-3-phenylpropanoic acid, sodium salt (Monomer Example Q) (1.71 grams of a 25% w/w solution in deionized water, 1.25 mmol) with S-BP (31.2 L of a 0.05 g/mL solution in deionized water). This mixture was diluted to a total of 5 grams with deionized water to produce a final monomer concentration of 0.25M and initiator concentration of 0.0312%. The procedure described in Example 1 was followed to provide a finished filter element with a graft density of 0.42 mmol/gram.

Example 25

[0313] A coating solution was prepared by mixing a solution of 4-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]butanoic acid, sodium salt (monomer Example M) (3.75 mL of a 0.25M solution in deionized water) with N,N-dimethylacrylamide (1.25 mL of a 0.25 M solution), and S-BP (31.25 L of a 0.05 g/mL solution in deionized water). The procedure described in Example 1 was followed to provide a finished filter element with a graft density of 0.14 mmol/gram.

Example 26

[0314] A coating solution was prepared by mixing a solution of 4-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]butanoic acid, sodium salt (monomer Example M) (2.5 mL of a 0.25M solution in deionized water) with N,N-dimethylacrylamide (2.5 mL of a 0.25 M solution), and S-BP (31.25 L of a 0.05 g/mL solution in deionized water). The procedure described in Example 1 was followed to provide a finished filter element with a graft density of 0.23 mmol/gram.

Example 27

[0315] A coating solution was prepared by mixing a solution of 4-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]butanoic acid, sodium salt (monomer Example M) (1.25 mL of a 0.25M solution in deionized water) with N,N-dimethylacrylamide (3.75 mL of a 0.25M solution), and S-BP (31.25 L of a 0.05 g/mL solution in deionized water). The procedure described in Example 1 was followed to provide a finished filter element with a graft density of 0.42 mmol/gram.

Example 28

[0316] A coating solution was prepared by mixing a solution of 4-[[2-methyl-2-(prop-2-enoylamino)propanoyl]amino]butanoic acid, sodium salt (monomer Example M) (1.25 mL of a 0.5M solution in deionized water) with N,N-dimethylacrylamide (3.75 mL of a 0.5M solution), and S-BP (31.25 L of a 0.05 g/mL solution in deionized water). The procedure described in Example 1 was followed to provide a finished filter element with a graft density of 0.43 mmol/gram.

Example 29

[0317] A coating solution was prepared by mixing a solution of 2-2-(2-methylprop-2-enoyloxy)ethylcarbamoylamino]ethanesulfonic acid, sodium salt (monomer Example I) (2.5 mL of a 0.75M solution in deionized water) with 3-phenyl-2-(prop-2-enoylamino)propanoic acid (2.5 mL of a 0.75M solution), and S-BP (500 L of a 0.05 g/mL solution in deionized water). The procedure described in Example 1 was followed to provide a finished filter element.

Example 30

[0318] A coating solution was prepared by mixing a solution of 2-[2-(2-methylprop-2-enoyloxy)ethylcarbamoylamino]ethanoic acid, sodium salt (monomer Example R) (1.62 grams of a 19.5% w/w solution in deionized water, 1.25 mmol) with S-BP (62.5 L of a 0.05 g/mL solution in deionized water). This mixture was diluted to a total of 5 grams with deionized water to produce a final monomer concentration of 0.25M and initiator concentration of 0.0625%. The procedure described in Example 1 was followed to provide a finished filter element with a graft density of 0.25 mmol/gram.

Example 31

Process for Separating Aggregated IgG from Monomeric IgG

[0319] The finished filter elements of Examples 1-30 were cut into 7.5 mm diameter disks. Two disks were inserted into each well of a 96-well Empore Filter Plate (Model 6065, 3M Corporation, St. Paul, Minn.) in which the original solid phase extraction material had been previously removed. The filter element disks were held in place with a plastic O-ring. The total working filter volume of each well was about 8.6 L. Each well was equilibrated with 1 mL of a non-IgG-containing buffer solution that had the same buffer composition as in the IgG-containing solution to be tested in the well. The equilibration solutions were removed from the wells using centrifugation (Allegra 25R centrifuge, Beckman Coulter, Brea, Calif.). The wells of the plate were then individually loaded (by pipet) with 1 mL of a single IgG-containing buffer solution selected from Table 2. The plate was sequentially centrifuged at 100, 200, 300, 600, 1200, and 3000 rcf (relative centrifugal force) or until the individual samples completely flowed through the filter elements. Each centrifugation step was conducted for about 5 minutes. The concentration of IgG in the IgG-containing buffer solution was either 3.32 mg/mL or 0.83 mg/mL, representing a challenge load of either 386 g/L or 96.5 g/L. Challenge load is defined as amount of IgG per given volume of filtration media (grams per liter, g/L). Separate collection plates were used for the equilibration solutions and IgG sample solutions. Filtered solutions were analyzed by size exclusion chromatography (SEC) using a Shimadzu Prominence HPLC (Shimadzu Scientific Instruments, Columbia, Md.) with a TSKgel G3000SWx1 column (Tosoh Bioscience LLC, Griesheim, Germany) (analysis conditions: injection volume of 20 L; flow rate of 1 ml/min; mobile phase: 100 mM sodium phosphate, 300 mM NaCl, pH 6.9; detection at 280 nanometers).

[0320] In Tables 3-35 the results are presented for the process of Example 31 using filter elements from Examples 1-30 and the IgG solutions (Number 1-27) described in Table 2. Monomer yield and aggregate removal were determined by comparison of peak areas of the starting and filtered solutions and the results are reported as the average of two replicates.

[0321] The percent of monomeric IgG present in the IgG solution before performing the separation process of Example 31 (Initial IgG solution) was determined using the SEC chromatography method described above. The peak area of the monomeric component (Peak A) of the sample was compared to the sum of the sample peak areas (Total Peak Area) according to Equation 1.

[0322] The percent of aggregated IgG present in the IgG solution before performing the separation process of Example 31 (Initial IgG solution) was determined using the SEC chromatography method described above. The peak area of the aggregate component (Peak B) of the sample was compared to the sum of the sample peak areas (Total Peak Area) according to Equation 2.

[0323] The ratio of the percent of monomeric IgG to the percent of aggregated IgG in the initial IgG solution (i.e. prior to performing the separation process of Example 31) was calculated according to Equation 3 and is reported as the Initial Composition Ratio.

[0324] The percent removal of aggregated IgG was determined using the SEC chromatography method described above and measuring the peak area for the aggregated component before performing the separation process (Peak B) and after performing the separation process of Example 31 (Peak C). The % Aggregated IgG Removed was calculated according to Equation 4.

[0325] The percent yield of monomeric IgG was determined using the SEC chromatography method described above and measuring the peak area for the monomeric component before performing the separation process (Peak A) and after performing the separation process of Example 31 (Peak D). The % Monomeric IgG Yield was calculated according to Equation 5.

[0326] The calculation for Monomeric IgG Recovered following the separation process of Example 31 was determined according to Equation 6 and is reported as Final Monomeric IgG Recovered (%).

[0327] The calculation for Aggregated IgG Recovered following the separation process of Example 31 was determined according to Equation 7 and is reported as Final Aggregated IgG Recovered (%).

[0328] The ratio of the percent of monomeric IgG to the percent of aggregated IgG in the final IgG solution (i.e. after performing the separation process of Example 31) was calculated according to Equation 8 and is reported as the Final Composition Ratio.

[0329] The Ratio of the Final Composition Ratio to the Initial Composition Ratio was calculated according to Equation 9.

[0330] The calculated values for Equations 1-9 are reported in Tables 3-35.

[00001]$\begin{array}{cc}\mathrm{Initial}.Math..Math.\mathrm{Monomeric}.Math..Math.\mathrm{IgG}.Math..Math.\left(\%\right)=\left(\frac{\mathrm{Peak}.Math..Math.\mathrm{Area}.Math..Math.A}{\mathrm{Total}.Math..Math.\mathrm{Peak}.Math..Math.\mathrm{Area}}\right)100& \mathrm{Equation}.Math..Math.1\\ \mathrm{Initial}.Math..Math.\mathrm{Aggregated}.Math..Math.\mathrm{IgG}.Math..Math.\left(\%\right)=\left(\frac{\mathrm{Peak}.Math..Math.\mathrm{Area}.Math..Math.B}{\mathrm{Total}.Math..Math.\mathrm{Peak}.Math..Math.\mathrm{Area}}\right)100& \mathrm{Equation}.Math..Math.2\\ \mathrm{Initial}.Math..Math.\mathrm{Composition}.Math..Math.\mathrm{Ratio}=\left(\frac{\mathrm{Initial}.Math..Math.\mathrm{Monomeric}.Math..Math.\mathrm{IgG}.Math..Math.\left(\%\right)}{\mathrm{Initial}.Math..Math.\mathrm{Aggregated}.Math..Math.\mathrm{IgG}.Math..Math.\left(\%\right)}\right)& \mathrm{Equation}.Math..Math.3\\ \mathrm{Aggregated}.Math..Math.\mathrm{IgG}.Math..Math.\mathrm{Removed}.Math..Math.\left(\%\right)=100-\left(\left(\frac{\mathrm{Peak}.Math..Math.\mathrm{Area}.Math..Math.C}{\mathrm{Peak}.Math..Math.\mathrm{Area}.Math..Math.B}\right)100\right)& \mathrm{Equation}.Math..Math.4\\ \mathrm{Monomeric}.Math..Math.\mathrm{IgG}.Math..Math.\mathrm{Yield}.Math..Math.\left(\%\right)=\left(\frac{\mathrm{Peak}.Math..Math.\mathrm{Area}.Math..Math.D}{\mathrm{Peak}.Math..Math.\mathrm{Area}.Math..Math.A}\right)100& \mathrm{Equation}.Math..Math.5\\ \mathrm{Final}.Math..Math.\mathrm{Monomeric}.Math..Math.\mathrm{IgG}.Math..Math.\mathrm{Recovered}.Math..Math.\left(\%\right)=\left(\frac{\begin{array}{c}\mathrm{Initial}.Math..Math.\mathrm{Monomeric}.Math..Math.\mathrm{IgG}.Math..Math.\left(\%\right)\\ \mathrm{Monomeric}.Math..Math.\mathrm{IgG}.Math..Math.\mathrm{Yield}.Math..Math.\left(\%\right)\end{array}}{100}\right)& \mathrm{Equation}.Math..Math.6\\ \mathrm{Final}.Math..Math.\mathrm{Aggregated}.Math..Math.\mathrm{IgG}.Math..Math.\mathrm{Recovered}.Math..Math.\left(\%\right)=\mathrm{Initial}.Math..Math.\mathrm{Aggregated}.Math..Math.\mathrm{IgG}.Math..Math.\left(\%\right).Math.\left(\frac{100-\mathrm{Aggregated}.Math..Math.\mathrm{IgG}.Math..Math.\mathrm{Removed}\left(\%\right)}{100}\right)& \mathrm{Equation}.Math..Math.7\\ \mathrm{Final}.Math..Math.\mathrm{Composition}.Math..Math.\mathrm{Ratio}.Math.=.Math.\left(\frac{\mathrm{Final}.Math..Math.\mathrm{Monomeric}.Math..Math.\mathrm{IgG}.Math..Math.\mathrm{Recovered}\left(\%\right)}{\mathrm{Final}.Math..Math.\mathrm{Aggregated}.Math..Math.\mathrm{IgG}.Math..Math.\mathrm{Recovered}.Math..Math.\left(\%\right)}\right)& \mathrm{Equation}.Math..Math.8\\ \mathrm{Ratio}.Math..Math.\mathrm{of}.Math..Math.\mathrm{Final}.Math..Math.\mathrm{Composition}.Math..Math.\mathrm{Ratio}.Math..Math.\mathrm{to}.Math..Math.\mathrm{Initial}.Math..Math.\mathrm{Composition}.Math..Math.\mathrm{Ratio}=\left(\frac{\mathrm{Final}.Math..Math.\mathrm{Composition}.Math..Math.\mathrm{Ratio}}{\mathrm{Initial}.Math..Math.\mathrm{Composition}.Math..Math.\mathrm{Ratio}}\right)& \mathrm{Equation}.Math..Math.9\end{array}$

TABLE-US-00003 TABLE 3 Filter Element of Example 1 386 g/L Challenge Load Filtration Performance Final Final Ratio of Final Initial Initial Aggregated Monomeric Monomeric Aggregated Composition IgG Monomeric Aggregated Initial IgG IgG IgG IgG Final Ratio to Initial Solution IgG IgG Composition Removed Yield Recovered Recovered Composition Composition Number (%) (%) Ratio (%) (%) (%) (%) Ratio Ratio 1 96.6 3.4 28.4 41.7 91.7 88.6 2.0 44.3 1.56 2 96.6 3.4 28.4 44.0 92.3 89.2 1.9 46.9 1.65 3 96.5 3.5 27.6 33.7 93.6 90.3 2.3 39.3 1.42 4 96.6 3.4 28.4 35.0 94.5 91.3 2.2 41.5 1.46 6 96.5 3.5 27.6 67.9 80.0 77.2 1.1 70.2 2.54 7 96.5 3.5 27.6 85.7 81.3 78.5 0.5 157.0 5.69 8 96.5 3.5 27.6 59.4 93.7 90.4 1.4 64.6 2.34 9 96.5 3.5 27.6 20.6 98.1 94.7 2.8 33.8 1.23 11 96.5 3.5 27.6 63.8 86.6 83.6 1.3 64.3 2.33 12 96.5 3.5 27.6 87.1 82.4 79.5 0.5 159.0 5.76 13 96.5 3.5 27.6 60.8 94.4 91.1 1.4 65.1 2.36 14 96.5 3.5 27.6 15.2 98.2 94.8 3.0 31.6 1.14 18 96.4 3.6 26.8 88.5 78.8 76.0 0.4 190.0 7.09 19 96.4 3.6 26.8 26.5 97.4 93.9 2.6 36.1 1.35 20 96.4 3.6 26.8 9.7 98.5 95.0 3.3 28.8 1.07 21 96.4 3.6 26.8 8.1 98.2 94.7 3.3 28.7 1.07 23 96.4 3.6 26.8 78.3 86.5 83.4 0.8 104.2 3.89 24 96.4 3.6 26.8 14.7 97.6 94.1 3.1 30.3 1.13 25 96.4 3.6 26.8 8.4 100.0 96.4 3.3 29.2 1.09

TABLE-US-00004 TABLE 4 Filter Element of Example 2 386 g/L Challenge Load Filtration Performance Final Final Ratio of Final Initial Initial Aggregated Monomeric Monomeric Aggregated Composition IgG Monomeric Aggregated Initial IgG IgG IgG IgG Final Ratio to Initial Solution IgG IgG Composition Removed Yield Recovered Recovered Composition Composition Number (%) (%) Ratio (%) (%) (%) (%) Ratio Ratio 1 96.6 3.4 28.4 89.6 87.6 84.6 0.4 211.5 7.45 2 96.5 3.5 27.6 73.0 95.5 92.2 0.9 102.4 3.71 3 96.5 3.5 27.6 54.4 97.3 93.9 1.6 58.7 2.13 4 96.6 3.4 28.4 9.8 98.1 94.8 3.1 30.6 1.08 6 96.5 3.5 27.6 78.7 89.5 86.4 0.7 123.4 4.47 7 96.5 3.5 27.6 44.7 97.6 94.2 1.9 49.6 1.80 8 96.5 3.5 27.6 16.9 98.3 94.9 2.9 32.7 1.18 9 96.5 3.5 27.6 7.9 98.4 95.0 3.2 29.7 1.08 11 96.6 3.4 28.4 68.8 87.4 84.4 1.1 76.7 2.70 12 96.5 3.5 27.6 49.6 97.3 93.9 1.8 52.2 1.89 13 96.5 3.5 27.6 35.8 98.0 94.6 2.2 43.0 1.56 14 96.5 3.5 27.6 9.7 98.3 94.9 3.2 29.6 1.07 18 96.5 3.5 27.6 80.4 95.6 92.3 0.7 131.8 4.77 19 96.5 3.5 27.6 17.9 98.3 94.9 2.9 32.7 1.18 20 96.5 3.5 27.6 5.2 98.8 95.3 3.3 28.9 1.05 21 96.5 3.5 27.6 4.1 98.6 95.1 3.4 28.0 1.01 23 96.5 3.5 27.6 54.7 93.8 90.5 1.6 56.6 2.05 24 96.4 3.6 26.8 5.3 98.7 95.1 3.4 28.0 1.04 25 96.5 3.5 27.6 4.0 98.9 95.4 3.4 28.0 1.01 26 96.5 3.5 27.6 6.2 99.0 95.5 3.3 28.9 1.05

TABLE-US-00005 TABLE 5 Filter Element of Example 3 386 g/L Challenge Load Filtration Performance Final Final Ratio of Final Initial Initial Aggregated Monomeric Monomeric Aggregated Composition IgG Monomeric Aggregated Initial IgG IgG IgG IgG Final Ratio to Initial Solution IgG IgG Composition Removed Yield Recovered Recovered Composition Composition Number (%) (%) Ratio (%) (%) (%) (%) Ratio Ratio 2 96.9 3.1 31.3 83.5 83.4 80.8 0.5 161.6 5.16 3 97.0 3.0 32.3 37.6 97.2 94.3 1.9 49.6 1.53 4 97.0 3.0 32.3 9.6 99.0 96.0 2.7 35.5 1.10 7 96.8 3.2 30.3 42.6 95.6 92.5 1.8 51.4 1.70 8 96.8 3.2 30.3 15.8 99.6 96.4 2.7 35.7 1.18 9 96.8 3.2 30.3 6.0 99.9 96.7 3.0 32.2 1.06 12 96.8 3.2 30.3 40.0 96.5 93.4 1.9 49.1 1.62 13 96.8 3.2 30.3 18.3 100.6 97.4 2.6 37.5 1.24 14 96.9 3.1 31.3 4.6 100.6 97.5 3.0 32.5 1.04 19 96.7 3.3 29.3 12.9 100.3 97.0 2.9 33.4 1.14 20 96.7 3.3 29.3 3.9 101.2 97.9 3.2 30.6 1.04 21 96.8 3.2 30.3 2.1 100.6 97.4 3.1 31.4 1.04

TABLE-US-00006 TABLE 6 Filter Element of Example 4 386 g/L Challenge Load Filtration Performance Final Final Ratio of Final Initial Initial Aggregated Monomeric Monomeric Aggregated Composition IgG Monomeric Aggregated Initial IgG IgG IgG IgG Final Ratio to Initial Solution IgG IgG Composition Removed Yield Recovered Recovered Composition Composition Number (%) (%) Ratio (%) (%) (%) (%) Ratio Ratio 1 97.3 2.7 36.0 11.5 98.5 95.8 2.4 39.9 1.11 2 96.6 3.4 28.4 46.7 93.2 90.0 1.8 50.0 1.76 3 96.7 3.3 29.3 7.5 98.1 94.9 3.1 30.6 1.04 4 96.7 3.3 29.3 5.5 98.2 95.0 3.1 30.6 1.04 6 96.6 3.4 28.4 23.9 97.4 94.1 2.6 36.2 1.28 7 96.5 3.5 27.6 7.5 98.7 95.2 3.2 29.7 1.08 8 96.5 3.5 27.6 5.7 98.4 95.0 3.3 28.8 1.04 9 96.5 3.5 27.6 3.9 98.6 95.1 3.4 28.0 1.01 11 96.4 3.6 26.8 57.4 96.1 92.6 1.5 61.7 2.30 12 96.5 3.5 27.6 15.9 97.9 94.5 2.9 32.6 1.18 13 96.5 3.5 27.6 7.7 98.6 95.1 3.2 29.7 1.08 14 96.5 3.5 27.6 5.9 98.1 94.7 3.3 28.7 1.04 18 96.5 3.5 27.6 36.1 96.8 93.4 2.2 42.4 1.51 19 96.3 3.7 26.0 6.3 98.2 94.6 3.5 27.0 1.04 20 96.3 3.7 26.0 3.3 98.9 95.2 3.6 26.4 1.02 21 96.3 3.7 26.0 3.1 99.0 95.3 3.6 26.5 1.02 23 96.4 3.6 26.8 60.2 90.6 87.3 1.4 62.3 2.32 24 96.3 3.7 26.0 4.5 98.9 95.2 3.5 27.2 1.05 25 96.3 3.7 26.0 2.9 99.0 95.3 3.6 26.5 1.02 26 96.3 3.7 26.0 2.5 99.0 95.3 3.6 26.5 1.02

TABLE-US-00007 TABLE 7 Filter Element of Example 4 96.5 g/L Challenge Load Filtration Performance Final Final Ratio of Final Initial Initial Aggregated Monomeric Monomeric Aggregated Composition IgG Monomeric Aggregated Initial IgG IgG IgG IgG Final Ratio to Initial Solution IgG IgG Composition Removed Yield Recovered Recovered Composition Composition Number (%) (%) Ratio (%) (%) (%) (%) Ratio Ratio 1 97.2 2.8 34.7 90.9 87.9 85.4 0.3 284.7 8.20 3 96.7 3.3 29.3 25.4 97.6 94.4 2.5 37.8 1.29 7 96.6 3.4 28.4 20.0 97.8 94.5 2.7 35.0 1.23 19 96.4 3.6 26.8 16.1 98.2 94.7 3.0 31.6 1.18 23 96.4 3.6 26.8 74.2 88.5 85.3 0.9 94.8 3.54 25 96.3 3.7 26.0 5.5 98.4 94.8 3.5 27.1 1.04

TABLE-US-00008 TABLE 8 Filter Element of Example 5 386 g/L Challenge Load Filtration Performance Final Final Ratio of Final Initial Initial Aggregated Monomeric Monomeric Aggregated Composition IgG Monomeric Aggregated Initial IgG IgG IgG IgG Final Ratio to Initial Solution IgG IgG Composition Removed Yield Recovered Recovered Composition Composition Number (%) (%) Ratio (%) (%) (%) (%) Ratio Ratio 1 97.0 3.0 32.3 9.4 97.1 94.2 2.7 34.9 1.08 2 96.7 3.3 29.3 14.1 96.7 93.5 2.8 33.4 1.14 3 96.7 3.3 29.3 15.9 96.2 93.0 2.8 33.2 1.13 6 96.6 3.4 28.4 65.9 83.6 80.8 1.2 67.3 2.37 7 96.6 3.4 28.4 80.1 85.8 82.9 0.7 118.4 4.16 8 96.5 3.5 27.6 43.7 95.5 92.2 2.0 46.1 1.67 11 96.5 3.5 27.6 59.0 87.3 84.2 1.4 60.1 2.17 12 96.5 3.5 27.6 82.8 85.1 82.1 0.6 136.8 4.96 13 96.5 3.5 27.6 44.7 96.0 92.6 1.9 48.7 1.76 18 96.5 3.5 27.6 89.8 82.7 79.8 0.4 199.5 7.23 19 96.4 3.6 26.8 21.0 98.3 94.8 2.8 33.8 1.26 20 96.4 3.6 26.8 9.4 98.7 95.1 3.3 28.8 1.07 23 96.3 3.7 26.0 75.7 81.8 78.8 0.9 87.6 3.37 24 96.3 3.7 26.0 11.4 98.5 94.9 3.3 28.8 1.11 25 96.3 3.7 26.0 7.7 99.0 95.3 3.4 28.0 1.08

TABLE-US-00009 TABLE 9 Filter Element of Example 6 386 g/L Challenge Load Filtration Performance Final Final Ratio of Final Initial Initial Aggregated Monomeric Monomeric Aggregated Composition IgG Monomeric Aggregated Initial IgG IgG IgG IgG Final Ratio to Initial Solution IgG IgG Composition Removed Yield Recovered Recovered Composition Composition Number (%) (%) Ratio (%) (%) (%) (%) Ratio Ratio 1 96.7 3.3 29.3 57.8 96.5 93.3 1.4 66.6 2.27 2 96.7 3.3 29.3 54.5 96.9 93.7 1.5 62.5 2.13 3 96.6 3.4 28.4 31.4 97.2 93.9 2.3 40.8 1.44 6 96.6 3.4 28.4 63.1 95.4 92.2 1.3 70.9 2.50 7 96.6 3.4 28.4 36.2 97.1 93.8 2.2 42.6 1.50 8 96.5 3.5 27.6 19.2 97.4 94.0 2.8 33.6 1.22 11 96.5 3.5 27.6 67.2 93.9 90.6 1.1 82.4 2.99 12 96.5 3.5 27.6 41.6 97.3 93.9 2.0 46.9 1.70 13 96.5 3.5 27.6 19.9 97.9 94.5 2.8 33.7 1.22 18 96.5 3.5 27.6 49.2 96.9 93.5 1.8 51.9 1.88 19 96.4 3.6 26.8 19.8 98.0 94.5 2.9 32.6 1.22 20 96.4 3.6 26.8 11.4 98.0 94.5 3.2 29.5 1.10

TABLE-US-00010 TABLE 10 Filter Element of Example 7 386 g/L Challenge Load Filtration Performance Final Final Ratio of Final Initial Initial Aggregated Monomeric Monomeric Aggregated Composition IgG Monomeric Aggregated Initial IgG IgG IgG IgG Final Ratio to Initial Solution IgG IgG Composition Removed Yield Recovered Recovered Composition Composition Number (%) (%) Ratio (%) (%) (%) (%) Ratio Ratio 1 96.5 3.5 27.6 76.8 84.8 81.8 0.8 102.2 3.70 2 96.7 3.3 29.3 91.4 88.4 85.5 0.3 285.0 9.72 3 96.7 3.3 29.3 47.2 97.2 94.0 1.7 55.3 1.89 6 96.7 3.3 29.3 86.0 76.3 73.8 0.5 147.6 5.04 7 96.6 3.4 28.4 51.8 96.5 93.2 1.6 58.2 2.05 8 96.6 3.4 28.4 19.6 98.5 95.2 2.7 35.2 1.24 11 96.5 3.5 27.6 74.6 77.2 74.5 0.9 82.8 3.00 12 96.5 3.5 27.6 63.9 95.4 92.1 1.3 70.8 2.56 13 96.6 3.4 28.4 17.4 98.9 95.5 2.8 34.1 1.20 18 96.5 3.5 27.6 89.2 89.3 86.2 0.4 215.5 7.81 19 96.4 3.6 26.8 17.7 98.8 95.2 3.0 31.7 1.18 20 96.4 3.6 26.8 7.2 99.2 95.6 3.3 29.0 1.08

TABLE-US-00011 TABLE 11 Filter Element of Example 8 386 g/L Challenge Load Filtration Performance Final Final Ratio of Final Initial Initial Aggregated Monomeric Monomeric Aggregated Composition IgG Monomeric Aggregated Initial IgG IgG IgG IgG Final Ratio to Initial Solution IgG IgG Composition Removed Yield Recovered Recovered Composition Composition Number (%) (%) Ratio (%) (%) (%) (%) Ratio Ratio 1 96.7 3.3 29.3 18.7 98.9 95.6 2.7 35.4 1.21 2 96.7 3.3 29.3 18.9 98.7 95.4 2.7 35.3 1.20 3 96.7 3.3 29.3 17.3 98.5 95.2 2.7 35.2 1.20 6 96.6 3.4 28.4 23.6 98.2 94.9 2.6 36.5 1.29 7 96.6 3.4 28.4 15.1 98.4 95.1 2.9 32.8 1.16 8 96.6 3.4 28.4 12.3 98.4 95.1 3.0 31.7 1.12 11 96.5 3.5 27.6 40.8 98.2 94.8 2.1 45.1 1.63 12 96.5 3.5 27.6 15.8 98.6 95.1 2.9 32.8 1.19 13 96.6 3.4 28.4 13.4 98.7 95.3 2.9 32.9 1.16 18 96.5 3.5 27.6 27.8 96.6 93.2 2.5 37.3 1.35 19 96.4 3.6 26.8 12.4 97.0 93.5 3.2 29.2 1.09 20 96.4 3.6 26.8 10.2 97.2 93.7 3.2 29.3 1.09 23 96.4 3.6 26.8 38.2 96.2 92.7 2.2 42.1 1.57 24 96.3 3.7 26.0 6.2 99.0 95.3 3.5 27.2 1.05 25 96.3 3.7 26.0 5.4 98.9 95.2 3.5 27.2 1.05

TABLE-US-00012 TABLE 12 Filter Element of Example 9 386 g/L Challenge Load Filtration Performance Final Final Ratio of Final Initial Initial Aggregated Monomeric Monomeric Aggregated Composition IgG Monomeric Aggregated Initial IgG IgG IgG IgG Final Ratio to Initial Solution IgG IgG Composition Removed Yield Recovered Recovered Composition Composition Number (%) (%) Ratio (%) (%) (%) (%) Ratio Ratio 1 96.7 3.3 29.3 67.1 95.5 92.3 1.1 83.9 2.86 2 96.6 3.4 28.4 65.0 96.3 93.0 1.2 77.5 2.73 3 96.6 3.4 28.4 35.3 97.6 94.3 2.2 42.9 1.51 6 96.6 3.4 28.4 73.3 91.5 88.4 0.9 98.2 3.46 7 96.6 3.4 28.4 40.9 97.4 94.1 2.0 47.0 1.65 8 96.5 3.5 27.6 17.0 98.2 94.8 2.9 32.7 1.18 11 96.5 3.5 27.6 70.0 88.7 85.6 1.1 77.8 2.82 12 96.5 3.5 27.6 49.9 97.2 93.8 1.8 52.1 1.89 13 96.5 3.5 27.6 18.3 98.7 95.2 2.9 32.8 1.19 15 96.6 3.4 28.4 82.6 88.1 85.1 0.6 142.7 5.02 16 96.4 3.6 26.8 31.1 98.3 94.8 2.5 37.9 1.41 17 96.4 3.6 26.8 13.8 98.8 95.2 3.1 30.7 1.15 18 96.4 3.6 26.8 65.4 96.2 92.7 1.2 77.2 2.88 19 96.3 3.7 26.0 19.4 98.8 95.1 3.0 31.7 1.22 20 96.3 3.7 26.0 11.4 98.8 95.1 3.3 28.8 1.11 23 96.3 3.7 26.0 46.2 95.3 91.8 2.0 45.9 1.77 24 96.2 3.8 25.3 8.4 98.7 94.9 3.5 27.1 1.07 25 96.2 3.8 25.3 5.3 98.9 95.1 3.6 26.4 1.04

TABLE-US-00013 TABLE 13 Filter Element of Example 10 386 g/L Challenge Load Filtration Performance Final Final Ratio of Final Initial Initial Aggregated Monomeric Monomeric Aggregated Composition IgG Monomeric Aggregated Initial IgG IgG IgG IgG Final Ratio to Initial Solution IgG IgG Composition Removed Yield Recovered Recovered Composition Composition Number (%) (%) Ratio (%) (%) (%) (%) Ratio Ratio 1 96.5 3.5 27.6 26.5 96.4 93.0 2.6 35.8 1.30 2 96.7 3.3 29.3 21.6 96.8 93.6 2.6 36.0 1.23 3 96.7 3.3 29.3 22.6 96.9 93.7 2.6 36.0 1.23 6 96.5 3.5 27.6 45.6 93.4 90.1 1.9 47.4 1.72 7 96.6 3.4 28.4 58.9 94.3 91.1 1.4 65.1 2.29 8 96.6 3.4 28.4 33.4 97.3 94.0 2.3 40.9 1.44 11 96.5 3.5 27.6 40.3 94.5 91.2 2.1 43.4 1.57 12 96.5 3.5 27.6 50.2 95.8 92.4 1.7 54.3 1.97 13 96.6 3.4 28.4 35.0 97.4 94.1 2.2 42.8 1.51 15 96.6 3.4 28.4 44.1 94.3 91.1 1.9 47.9 1.69 16 96.4 3.6 26.8 46.6 97.1 93.6 1.9 49.3 1.84 17 96.3 3.7 26.0 24.9 98.3 94.7 2.8 33.8 1.30 18 96.5 3.5 27.6 81.3 92.6 89.4 0.7 127.7 4.63 19 96.4 3.6 26.8 19.9 98.9 95.3 2.9 32.9 1.23 20 96.4 3.6 26.8 11.5 99.0 95.4 3.2 29.8 1.11 23 96.4 3.6 26.8 53.4 93.7 90.3 1.7 53.1 1.98 24 96.3 3.7 26.0 14.1 98.6 95.0 3.2 29.7 1.14 25 96.3 3.7 26.0 11.3 98.8 95.1 3.3 28.8 1.11

TABLE-US-00014 TABLE 14 Filter Element of Example 11 386 g/L Challenge Load Filtration Performance Final Final Ratio of Final Initial Initial Aggregated Monomeric Monomeric Aggregated Composition IgG Monomeric Aggregated Initial IgG IgG IgG IgG Final Ratio to Initial Solution IgG IgG Composition Removed Yield Recovered Recovered Composition Composition Number (%) (%) Ratio (%) (%) (%) (%) Ratio Ratio 1 96.7 3.3 29.3 61.1 90.9 87.9 1.3 67.6 2.31 2 96.7 3.3 29.3 77.4 93.2 90.1 0.7 128.7 4.39 3 96.7 3.3 29.3 35.7 97.7 94.5 2.1 45.0 1.54 6 96.7 3.3 29.3 80.7 85.9 83.1 0.6 138.5 4.72 7 96.6 3.4 28.4 45.5 97.0 93.7 1.9 49.3 1.74 8 96.6 3.4 28.4 36.5 97.8 94.5 2.2 43.0 1.51 11 96.6 3.4 28.4 65.4 89.7 86.7 1.2 72.2 2.54 12 96.6 3.4 28.4 44.3 97.4 94.1 1.9 49.5 1.74 13 96.6 3.4 28.4 18.3 98.5 95.2 2.8 34.0 1.20 15 96.8 3.2 30.3 66.6 88.3 85.5 1.1 77.7 2.56 16 96.5 3.5 27.6 30.7 98.2 94.8 2.4 39.5 1.43 17 96.5 3.5 27.6 14.1 98.9 95.4 3.0 31.8 1.15 18 96.5 3.5 27.6 78.4 93.9 90.6 0.8 113.2 4.10 19 96.4 3.6 26.8 17.9 98.9 95.3 3.0 31.8 1.19 20 96.4 3.6 26.8 9.6 98.9 95.3 3.3 28.9 1.08

TABLE-US-00015 TABLE 15 Filter Element of Example 12 387 g/L Challenge Load Filtration Performance Final Final Ratio of Final Initial Initial Aggregated Monomeric Monomeric Aggregated Composition IgG Monomeric Aggregated Initial IgG IgG IgG IgG Final Ratio to Initial Solution IgG IgG Composition Removed Yield Recovered Recovered Composition Composition Number (%) (%) Ratio (%) (%) (%) (%) Ratio Ratio 1 96.8 3.2 30.3 45.4 96.6 93.5 1.7 55.0 1.82 2 96.7 3.3 29.3 30.5 98.4 95.2 2.3 41.4 1.41 3 96.7 3.3 29.3 13.3 99.4 96.1 2.9 33.1 1.13 4 96.7 3.3 29.3 2.4 100.3 97.0 3.2 30.3 1.03 6 96.7 3.3 29.3 55.3 96.6 93.4 1.5 62.2 2.12 7 96.7 3.3 29.3 14.6 100.1 96.8 2.8 34.6 1.18 8 96.6 3.4 28.4 2.8 101.0 97.6 3.3 29.6 1.04 9 96.6 3.4 28.4 0.3 100.3 96.9 3.4 28.5 1.00 18 96.4 3.6 26.8 29.5 98.2 94.7 2.5 37.9 1.41 19 96.3 3.7 26.0 2.7 99.5 95.8 3.6 26.6 0.99 20 96.3 3.7 26.0 1.2 100.8 97.1 3.7 26.2 1.01 21 96.3 3.7 26.0 0.7 100.8 97.1 3.7 26.2 1.01 23 96.3 3.7 26.0 10.9 99.3 95.6 3.3 29.0 1.11 24 96.2 3.8 25.3 0.5 99.5 95.7 3.8 25.2 1.00 25 96.2 3.8 25.3 0.3 100.1 96.3 3.8 25.3 1.00 26 96.2 3.8 25.3 0.0 100.2 96.4 3.8 25.4 1.00

TABLE-US-00016 TABLE 16 Filter Element of Example 12 96.5 g/L Challenge Load Filtration Performance Final Final Ratio of Final Initial Initial Aggregated Monomeric Monomeric Aggregated Composition IgG Monomeric Aggregated Initial IgG IgG IgG IgG Final Ratio to Initial Solution IgG IgG Composition Removed Yield Recovered Recovered Composition Composition Number (%) (%) Ratio (%) (%) (%) (%) Ratio Ratio 1 97.2 2.8 34.7 83.8 87.6 85.1 0.5 170.2 4.90 3 96.7 3.3 29.3 25.5 97.9 94.7 2.5 37.9 1.29 7 96.6 3.4 28.4 15.4 98.3 95.0 2.9 32.8 1.15 19 96.4 3.6 26.8 5.4 98.8 95.2 3.4 28.0 1.04 23 96.4 3.6 26.8 20.7 97.0 93.5 2.9 32.2 1.20 25 96.3 3.7 26.0 2.9 99.2 95.5 3.6 26.5 1.02

TABLE-US-00017 TABLE 17 Filter Element of Example 13 387 g/L Challenge Load Filtration Performance Final Final Ratio of Final Initial Initial Aggregated Monomeric Monomeric Aggregated Composition IgG Monomeric Aggregated Initial IgG IgG IgG IgG Final Ratio to Initial Solution IgG IgG Composition Removed Yield Recovered Recovered Composition Composition Number (%) (%) Ratio (%) (%) (%) (%) Ratio Ratio 1 96.8 3.2 30.3 28.5 98.2 95.1 2.3 41.3 1.36 2 96.7 3.3 29.3 18.5 98.6 95.3 2.7 35.3 1.20 3 96.7 3.3 29.3 11.9 99.1 95.8 2.9 33.0 1.13 4 96.7 3.3 29.3 8.3 99.6 96.3 3.0 32.1 1.09 6 96.7 3.3 29.3 32.3 98.4 95.2 2.2 43.3 1.48 7 96.7 3.3 29.3 12.1 99.9 96.6 2.9 33.3 1.14 8 96.6 3.4 28.4 7.0 100.6 97.2 3.2 30.4 1.07 9 96.6 3.4 28.4 5.3 99.9 96.5 3.2 30.2 1.06 18 96.4 3.6 26.8 22.2 98.3 94.8 2.8 33.8 1.26 19 96.3 3.7 26.0 5.9 99.1 95.4 3.5 27.3 1.05 20 96.3 3.7 26.0 4.6 100.4 96.7 3.5 27.6 1.06 21 96.3 3.7 26.0 4.4 100.4 96.7 3.5 27.6 1.06 23 96.3 3.7 26.0 11.7 99.0 95.3 3.3 28.9 1.16 24 96.2 3.8 25.3 3.4 99.5 95.7 3.7 25.9 1.02 25 96.2 3.8 25.3 2.7 100.0 96.2 3.7 26.0 1.03 26 96.2 3.8 25.3 2.4 100.1 96.3 3.7 26.0 1.03

TABLE-US-00018 TABLE 18 Filter Element of Example 13 96.5 g/L Challenge Load Filtration Performance Final Final Ratio of Final Initial Initial Aggregated Monomeric Monomeric Aggregated Composition IgG Monomeric Aggregated Initial IgG IgG IgG IgG Final Ratio to Initial Solution IgG IgG Composition Removed Yield Recovered Recovered Composition Composition Number (%) (%) Ratio (%) (%) (%) (%) Ratio Ratio 1 97.2 2.8 34.7 54.4 94.3 91.7 1.3 70.5 2.03 3 96.7 3.3 29.3 41.1 96.8 93.6 1.9 49.3 1.68 7 96.6 3.4 28.4 34.4 97.0 93.7 2.2 42.6 1.50 19 96.4 3.6 26.8 18.6 97.9 94.4 2.9 32.5 1.21 23 96.4 3.6 26.8 32.4 95.5 92.1 2.4 38.4 1.43 25 96.3 3.7 26.0 11.3 98.2 94.6 3.3 28.7 1.10

TABLE-US-00019 TABLE 19 Filter Element of Example 14 386 g/L Challenge Load Filtration Performance Final Final Ratio of Final Initial Initial Aggregated Monomeric Monomeric Aggregated Composition IgG Monomeric Aggregated Initial IgG IgG IgG IgG Final Ratio to Initial Solution IgG IgG Composition Removed Yield Recovered Recovered Composition Composition Number (%) (%) Ratio (%) (%) (%) (%) Ratio Ratio 1 97.2 2.8 34.7 61.4 73.0 71.0 1.1 64.5 1.86 2 96.5 3.5 27.6 41.0 96.7 93.3 2.1 44.4 1.61 3 96.8 3.2 30.3 5.9 99.0 95.8 3.0 31.9 1.05 4 96.8 3.2 30.3 5.9 99.0 95.8 3.0 31.9 1.05 5 96.5 3.5 27.6 1.2 98.9 95.4 3.5 27.2 0.99 6 96.6 3.4 28.4 81.3 71.2 68.8 0.6 114.7 4.04 7 96.5 3.5 27.6 53.4 95.6 92.3 1.6 57.7 2.09 8 96.5 3.5 27.6 7.8 98.5 95.1 3.2 29.7 1.08 9 96.7 3.3 29.3 0.3 99.9 96.6 3.3 29.3 1.00 10 96.4 3.6 26.8 0.0 99.7 96.1 3.6 26.7 1.00 18 96.4 3.6 26.8 80.4 81.4 78.5 0.7 112.1 4.18 19 96.2 3.8 25.3 9.0 98.9 95.1 3.5 27.2 1.08 20 96.3 3.7 26.0 2.2 99.1 95.4 3.6 26.5 1.02 21 96.3 3.7 26.0 1.3 99.0 95.3 3.7 25.8 0.99 22 96.3 3.7 26.0 0.5 99.8 96.1 3.7 26.0 1.00 23 96.4 3.6 26.8 67.0 85.3 82.2 1.2 68.5 2.56 24 96.6 3.4 28.4 3.3 98.4 95.1 3.3 28.8 1.01 25 96.3 3.7 26.0 3.9 98.6 95.0 3.6 26.4 1.01 26 96.2 3.8 25.3 3.3 98.7 94.9 3.7 25.6 1.01 27 96.2 3.8 25.3 1.6 98.6 94.9 3.7 25.6 1.01

TABLE-US-00020 TABLE 20 Filter Element of Example 15 386 g/L Challenge Load Filtration Performance Final Final Ratio of Final Initial Initial Aggregated Monomeric Monomeric Aggregated Composition IgG Monomeric Aggregated Initial IgG IgG IgG IgG Final Ratio to Initial Solution IgG IgG Composition Removed Yield Recovered Recovered Composition Composition Number (%) (%) Ratio (%) (%) (%) (%) Ratio Ratio 1 97.2 2.8 34.7 73.8 68.0 66.1 0.7 94.41 2.72 2 96.5 3.5 27.6 58.7 97.5 94.1 1.4 67.2 2.43 3 96.8 3.2 30.3 2.6 99.1 95.9 3.1 30.9 1.02 4 96.8 3.2 30.3 2.6 99.1 95.9 3.1 30.9 1.02 5 96.5 3.5 27.6 0.6 98.8 95.3 3.5 27.2 0.99 6 96.6 3.4 28.4 52.8 75.9 73.3 1.6 45.8 1.61 7 96.5 3.5 27.6 33.4 97.1 93.7 2.3 40.7 1.46 8 96.5 3.5 27.6 6.8 99.0 95.5 3.3 28.9 1.05 9 96.7 3.3 29.3 0.7 99.9 96.6 3.3 29.3 1.00 10 96.4 3.6 26.8 0.6 99.8 96.2 3.6 26.7 1.00 18 96.4 3.6 26.8 89.9 83.9 80.9 0.4 202.2 7.54 19 96.2 3.8 25.3 3.3 99.9 96.1 3.7 26.0 1.03 20 96.3 3.7 26.0 0.9 99.9 96.2 3.7 26.0 1.00 21 96.3 3.7 26.0 1.1 99.3 95.6 3.7 25.8 0.99 22 96.3 3.7 26.0 0.2 100.2 96.5 3.7 26.1 1.00 23 96.4 3.6 26.8 56.8 86.1 83.0 1.6 51.9 1.94 24 96.6 3.4 28.4 7.5 98.7 95.3 3.1 30.7 1.08 25 96.3 3.7 26.0 1.5 99.3 95.6 3.6 26.5 1.02 26 96.2 3.8 25.3 1.8 99.1 95.3 3.7 25.8 1.02 27 96.2 3.8 25.3 2.1 99.0 95.2 3.7 25.7 1.02

TABLE-US-00021 TABLE 21 Filter Element of Example 16 386 g/L Challenge Load Filtration Performance Final Final Ratio of Final Initial Initial Aggregated Monomeric Monomeric Aggregated Composition IgG Monomeric Aggregated Initial IgG IgG IgG IgG Final Ratio to Initial Solution IgG IgG Composition Removed Yield Recovered Recovered Composition Composition Number (%) (%) Ratio (%) (%) (%) (%) Ratio Ratio 1 97.2 2.8 34.7 72.0 87.1 84.7 0.8 105.9 3.05 2 96.5 3.5 27.6 70.2 91.9 88.7 1.0 88.7 3.21 3 96.8 3.2 30.3 28.6 98.7 95.5 2.3 41.5 1.37 4 96.8 3.2 30.3 28.6 98.7 95.5 2.3 41.5 1.37 5 96.5 3.5 27.6 1.1 99.3 95.8 3.5 27.4 0.99 6 96.6 3.4 28.4 77.4 79.2 76.5 0.8 95.6 3.37 7 96.5 3.5 27.6 68.7 93.6 90.3 1.1 82.1 2.97 8 96.5 3.5 27.6 24.5 99.2 95.7 2.6 36.8 1.33 9 96.7 3.3 29.3 12.1 100.1 96.8 2.9 33.4 1.14 18 96.4 3.6 26.8 86.1 88.7 85.5 0.5 171.0 6.38 19 96.2 3.8 25.3 4.9 100.5 96.7 3.6 26.9 1.06 21 96.3 3.7 26.0 0.3 99.8 96.1 3.7 26.0 1.00 23 96.4 3.6 26.8 55.7 91.2 87.9 1.6 54.9 2.05 24 96.6 3.4 28.4 2.2 99.4 96.0 3.3 29.1 1.02 25 96.3 3.7 26.0 0.7 99.2 95.5 3.7 25.8 0.99 26 96.2 3.8 25.3 1.7 99.0 95.2 3.7 257 1.01 27 96.2 3.8 25.3 1.0 99.7 95.9 3.8 25.2 1.00

TABLE-US-00022 TABLE 22 Filter Element of Example 17 386 g/L Challenge Load Filtration Performance Final Final Ratio of Final Initial Initial Aggregated Monomeric Monomeric Aggregated Composition IgG Monomeric Aggregated Initial IgG IgG IgG IgG Final Ratio to Initial Solution IgG IgG Composition Removed Yield Recovered Recovered Composition Composition Number (%) (%) Ratio (%) (%) (%) (%) Ratio Ratio 1 97.2 2.8 34.7 97.5 63.6 61.8 0.1 618.0 17.80 2 96.5 3.5 27.6 33.5 97.4 94.0 2.3 40.9 1.48 3 96.8 3.2 30.3 1.3 99.9 96.7 3.2 30.2 1.00 4 96.8 3.2 30.3 1.3 99.9 96.7 3.2 30.2 1.00 5 96.5 3.5 27.6 0.2 99.7 96.2 3.5 27.5 1.00 6 96.6 3.4 28.4 95.3 66.8 64.5 0.2 322.5 11.4 7 96.5 3.5 27.6 37.9 97.7 94.3 2.2 42.9 1.55 8 96.5 3.5 27.6 15.9 88.6 85.5 2.9 29.5 1.07 18 96.4 3.6 26.8 64.8 82.4 79.4 1.3 61.1 2.28 19 96.2 3.8 25.3 0.9 100.8 97.0 3.8 25.5 1.01 21 96.3 3.7 26.0 0.0 100.1 96.4 3.7 26.0 1.00 24 96.6 3.4 28.4 1.3 99.5 96.1 3.4 28.3 1.00 25 96.3 3.7 26.0 0.5 99.9 96.2 3.7 26.0 1.00 26 96.2 3.8 25.3 1.1 99.4 95.6 3.8 25.2 1.00 27 96.2 3.8 25.3 97.5 63.6 61.2 0.1 612.0 24.19

TABLE-US-00023 TABLE 23 Filter Element of Example 18 387 g/L Challenge Load Filtration Performance Final Final Ratio of Final Initial Initial Aggregated Monomeric Monomeric Aggregated Composition IgG Monomeric Aggregated Initial IgG IgG IgG IgG Final Ratio to Initial Solution IgG IgG Composition Removed Yield Recovered Recovered Composition Composition Number (%) (%) Ratio (%) (%) (%) (%) Ratio Ratio 1 96.8 3.2 30.3 95.0 83.3 80.6 0.2 403.0 13.30 2 96.7 3.3 29.3 92.5 87.7 84.8 0.2 424.0 14.47 3 96.7 3.3 29.3 42.3 97.3 94.1 1.9 49.5 1.69 4 96.6 3.4 28.4 3.0 99.9 96.5 3.3 29.2 1.03 6 96.6 3.4 28.4 94.8 75.0 72.5 0.2 362.5 12.76 7 96.7 3.3 29.3 48.3 96.7 93.5 1.7 55.0 1.87 8 96.6 3.4 28.4 2.2 99.9 96.5 3.3 29.2 1.02 9 96.5 3.5 27.6 0.2 101.1 97.6 3.5 27.9 1.01 18 96.3 3.7 26.0 92.5 86.7 83.5 0.3 278.3 10.70 19 96.3 3.7 26.0 2.2 100.6 96.9 3.6 26.9 1.03 20 96.3 3.7 26.0 0.7 100.3 96.6 3.7 26.1 1.00 21 96.3 3.7 26.0 0.6 100.4 96.7 3.7 26.1 1.00 23 96.2 3.8 25.3 63.8 90.5 87.1 1.4 62.2 2.46

TABLE-US-00024 TABLE 24 Filter Element of Example 19 387 g/L Challenge Load Filtration Performance Final Final Ratio of Final Initial Initial Aggregated Monomeric Monomeric Aggregated Composition IgG Monomeric Aggregated Initial IgG IgG IgG IgG Final Ratio to Initial Solution IgG IgG Composition Removed Yield Recovered Recovered Composition Composition Number (%) (%) Ratio (%) (%) (%) (%) Ratio Ratio 1 96.8 3.2 30.3 90.7 86.1 83.3 0.3 277.7 9.17 2 96.7 3.3 29.3 91.6 87.7 84.8 0.3 282.7 9.65 3 96.7 3.3 29.3 66.6 94.5 91.4 1.1 83.1 2.83 4 96.6 3.4 28.4 15.0 99.0 95.6 2.9 33.0 1.16 6 96.6 3.4 28.4 93.5 75.2 72.6 0.2 363.0 12.78 7 96.7 3.3 29.3 54.8 95.5 92.3 1.5 61.5 2.10 8 96.6 3.4 28.4 7.8 99.2 95.8 3.1 30.9 1.09 9 96.5 3.5 27.6 1.8 100.6 97.1 3.4 28.5 1.03 18 96.3 3.7 26.0 94.7 83.6 80.5 0.2 402.5 15.48 19 96.3 3.7 26.0 5.8 100.3 96.6 3.5 27.6 1.06 20 96.3 3.7 26.0 1.9 100.4 96.7 3.6 26.9 1.03 21 96.3 3.7 26.0 1.0 100.3 96.6 3.7 26.1 1.00 23 96.2 3.8 25.3 70.1 86.9 83.6 1.1 76.0 3.00 25 96.2 3.8 25.3 0.3 101.1 97.3 3.8 25.6 1.01

TABLE-US-00025 TABLE 25 Filter Element of Example 20 387 g/L Challenge Load Filtration Performance Final Final Ratio of Final Initial Initial Aggregated Monomeric Monomeric Aggregated Composition IgG Monomeric Aggregated Initial IgG IgG IgG IgG Final Ratio to Initial Solution IgG IgG Composition Removed Yield Recovered Recovered Composition Composition Number (%) (%) Ratio (%) (%) (%) (%) Ratio Ratio 1 96.8 3.2 30.3 57.6 93.3 90.3 1.4 64.5 2.13 2 96.7 3.3 29.3 76.9 89.8 86.8 0.8 108.5 3.70 3 96.7 3.3 29.3 75.9 90.7 87.7 0.8 109.6 3.74 4 96.6 3.4 28.4 54.5 95.8 92.5 1.5 61.7 2.17 6 96.6 3.4 28.4 79.9 79.6 76.9 0.7 109.9 3.87 7 96.7 3.3 29.3 81.1 89.2 86.3 0.6 143.8 4.91 8 96.6 3.4 28.4 12.1 98.7 95.3 3.0 31.8 1.12 9 96.5 3.5 27.6 5.8 99.5 96.0 3.3 29.1 1.06 18 96.3 3.7 26.0 76.9 84.0 80.9 0.9 89.9 3.46 19 96.3 3.7 26.0 8.5 99.1 95.4 3.4 28.1 1.08 20 96.3 3.7 26.0 4.6 99.2 95.5 3.5 27.3 1.05 21 96.3 3.7 26.0 3.8 99.5 95.8 3.6 26.6 1.02 23 96.2 3.8 25.3 50.7 89.8 86.4 1.9 45.5 1.80 24 96.1 3.9 24.6 5.0 99.2 95.3 3.7 25.7 1.04 25 96.2 3.8 25.3 3.1 99.4 95.6 3.7 25.8 1.02 26 96.2 3.8 25.3 4.1 99.4 95.6 3.6 26.5 1.05

TABLE-US-00026 TABLE 26 Filter Element of Example 21 387 g/L Challenge Load Filtration Performance Final Final Ratio of Final Initial Initial Aggregated Monomeric Monomeric Aggregated Composition IgG Monomeric Aggregated Initial IgG IgG IgG IgG Final Ratio to Initial Solution IgG IgG Composition Removed Yield Recovered Recovered Composition Composition Number (%) (%) Ratio (%) (%) (%) (%) Ratio Ratio 1 96.8 3.2 30.1 3.4 97.1 94.0 3.1 30.2 1.0 2 96.7 3.3 29.4 6.4 96.6 93.4 3.1 30.3 1.0 3 96.7 3.3 29.2 5.6 96.8 93.6 3.1 29.9 1.0 4 96.6 3.4 28.8 6.9 96.8 93.6 3.1 29.9 1.0 6 96.6 3.4 28.2 68.1 71.2 68.8 1.1 63.0 2.2 7 96.7 3.3 28.9 87.7 76.7 74.2 0.4 180.4 6.2 8 96.6 3.4 28.0 40.7 96.6 93.2 2.0 45.6 1.6 9 96.5 3.5 27.9 8.4 99.8 96.4 3.2 30.4 1.1 18 96.3 3.7 26.3 88.1 69.8 67.2 0.4 154.7 5.9 19 96.3 3.7 25.9 7.7 100.2 96.5 3.4 28.1 1.1 20 96.3 3.7 26.2 0.4 100.2 96.6 3.7 26.4 1.0 21 96.3 3.7 26.3 1.0 100.3 96.6 3.6 26.6 1.0 23 96.2 3.8 25.3 68.4 82.4 79.3 1.2 66.0 2.6 24 96.1 3.9 24.9 1.9 100.1 96.2 3.8 25.4 1.0 25 96.2 3.8 25.1 1.7 99.9 96.0 3.8 25.5 1.0 26 96.2 3.8 25.2 0.3 100.6 96.8 3.8 25.3 1.0

TABLE-US-00027 TABLE 27 Filter Element of Example 22 387 g/L Challenge Load Filtration Performance Final Final Ratio of Final Initial Initial Aggregated Monomeric Monomeric Aggregated Composition IgG Monomeric Aggregated Initial IgG IgG IgG IgG Final Ratio to Initial Solution IgG IgG Composition Removed Yield Recovered Recovered Composition Composition Number (%) (%) Ratio (%) (%) (%) (%) Ratio Ratio 1 96.8 3.2 30.3 2.9 96.5 93.4 3.1 30.1 0.99 2 96.7 3.3 29.3 6.1 96.0 92.8 3.1 29.9 1.02 3 96.7 3.3 29.3 11.1 96.2 93.0 2.9 32.1 1.09 4 96.6 3.4 28.4 6.6 96.7 93.4 3.2 29.2 1.03 6 96.6 3.4 28.4 73.3 73.3 70.8 0.9 78.7 2.77 7 96.7 3.3 29.3 82.4 76.8 74.3 0.6 123.8 4.23 8 96.6 3.4 28.4 33.3 97.9 94.6 2.3 41.1 1.45 9 96.5 3.5 27.6 8.7 99.6 96.1 3.2 30.0 1.09 18 96.3 3.7 26.0 79.6 80.3 77.3 0.8 96.6 3.71 19 96.3 3.7 26.0 5.1 99.8 96.1 3.5 27.4 1.05 20 96.3 3.7 26.0 0.4 100.4 96.7 3.7 26.2 1.01 21 96.3 3.7 26.0 0.8 100.3 96.6 3.7 26.1 1.00 23 96.2 3.8 25.3 51.4 86.6 83.3 1.8 46.3 1.83 24 96.1 3.9 24.6 1.0 100.2 96.3 3.9 24.7 1.00 25 96.2 3.8 25.3 1.7 99.6 95.8 3.7 25.9 1.02 26 96.2 3.8 25.3 0.2 100.5 96.7 3.8 25.4 1.00

TABLE-US-00028 TABLE 28 Filter Element of Example 23 386 g/L Challenge Load Filtration Performance Final Final Ratio of Final Initial Initial Aggregated Monomeric Monomeric Aggregated Composition IgG Monomeric Aggregated Initial IgG IgG IgG IgG Final Ratio to Initial Solution IgG IgG Composition Removed Yield Recovered Recovered Composition Composition Number (%) (%) Ratio (%) (%) (%) (%) Ratio Ratio 1 97.5 2.5 39.0 10.8 91.9 89.6 2.2 40.7 1.04 2 97.6 2.4 40.7 5.9 93.3 91.1 2.3 39.6 0.97 3 97.4 2.6 37.5 21.1 94.2 91.8 2.1 43.7 1.17 4 97.3 2.7 36.0 33.3 95.1 92.5 1.8 51.4 1.43 5 97.4 2.6 37.5 29.9 97.2 94.7 1.8 52.6 1.40 6 97.2 2.8 34.7 17.2 92.2 89.6 2.3 38.9 1.12 7 97.0 3.0 32.3 38.3 91.0 88.3 1.9 46.5 1.44 8 97.0 3.0 32.3 46.8 93.9 91.1 1.6 56.9 1.76 9 97.1 2.9 33.5 43.0 96.4 93.6 1.7 55.0 1.64 10 97.4 2.6 37.5 21.2 98.4 95.8 2.0 47.9 1.28 18 96.8 3.2 30.3 57.4 87.2 84.4 1.4 60.3 1.99 19 96.6 3.4 28.4 69.0 93.3 90.1 1.1 81.9 2.88 20 96.8 3.2 30.3 41.5 97.9 94.8 1.9 49.9 1.65 21 96.9 3.1 31.3 21.9 98.5 95.4 2.4 39.8 1.27 22 97.3 2.7 36.0 10.3 98.9 96.2 2.4 40.1 1.11 23 96.7 3.3 29.3 86.4 81.4 78.7 0.4 196.8 6.72 24 96.7 3.3 29.3 43.3 97.4 94.2 1.9 49.6 1.69 25 96.7 3.3 29.3 21.0 98.8 95.5 2.6 36.7 1.25 26 96.7 3.3 29.3 10.0 99.2 95.9 3.0 32.0 1.10 27 97.2 2.8 34.7 5.5 98.9 96.1 2.6 37.0 1.07

TABLE-US-00029 TABLE 29 Filter Element of Example 24 387 g/L Challenge Load Filtration Performance Final Final Ratio of Final Initial Initial Aggregated Monomeric Monomeric Aggregated Composition IgG Monomeric Aggregated Initial IgG IgG IgG IgG Final Ratio to Initial Solution IgG IgG Composition Removed Yield Recovered Recovered Composition Composition Number (%) (%) Ratio (%) (%) (%) (%) Ratio Ratio 1 96.4 3.6 26.8 10.7 92.7 89.4 3.2 27.9 1.04 2 97.1 2.9 33.5 8.0 92.5 89.8 2.7 33.2 0.99 3 96.7 3.3 29.3 14.5 93.8 90.7 2.8 32.4 1.10 4 96.9 3.1 31.3 19.2 93.3 90.4 2.5 36.1 1.15 6 96.8 3.2 30.3 22.1 93.1 90.1 2.5 36.0 1.19 7 96.5 3.5 27.6 48.5 83.9 81.0 1.8 45.0 1.63 8 95.3 4.7 20.3 66.7 76.6 73.0 1.6 45.6 2.25 9 97.3 2.7 36.0 71.4 80.1 77.9 0.8 97.4 2.71 18 95.6 4.4 21.7 71.9 63.0 60.2 1.2 50.2 2.31 19 94.9 5.1 18.6 75.0 86.3 81.9 1.3 63.0 3.39 20 97.1 2.9 33.5 36.3 95.4 92.6 1.8 51.4 1.53 21 96.5 3.5 27.6 17.7 98.7 95.2 2.9 32.8 1.19 23 94.7 5.3 17.9 81.8 68.5 64.9 1.0 64.9 3.63 24 94.6 5.4 17.5 22.6 97.1 91.9 4.2 21.9 1.25 25 94.7 5.3 17.9 7.6 99.5 94.2 4.9 19.2 1.08 26 96.1 3.9 24.6 6.5 99.9 96.0 3.6 26.7 1.08

TABLE-US-00030 TABLE 30 Filter Element of Example 25 386 g/L Challenge Load Filtration Performance Final Final Ratio of Final Initial Initial Aggregated Monomeric Monomeric Aggregated Composition IgG Monomeric Aggregated Initial IgG IgG IgG IgG Final Ratio to Initial Solution IgG IgG Composition Removed Yield Recovered Recovered Composition Composition Number (%) (%) Ratio (%) (%) (%) (%) Ratio Ratio 1 97.1 2.9 33.5 94.8 86.5 84.0 0.2 420.0 12.54 2 97.1 2.9 33.5 83.0 91.3 88.7 0.5 177.4 5.30 3 96.6 3.4 28.4 26.1 97.4 94.1 2.5 37.6 1.32 6 96.8 3.2 30.3 93.1 79.1 76.6 0.2 383.0 12.64 7 96.5 3.5 27.6 32.6 97.1 93.7 2.4 39.0 1.41 8 98.4 1.6 61.5 6.4 98.1 96.5 1.5 64.3 1.05 18 96.0 4.0 24.0 79.8 87.4 83.9 0.8 104.9 4.37 19 96.2 3.8 25.3 8.9 98.5 94.8 3.5 27.1 1.07 20 96.2 3.8 25.3 5.2 99.6 95.8 3.6 26.6 1.05 23 96.2 3.8 25.3 50.8 88.6 85.2 1.9 44.8 1.77 24 96.0 4.0 24.0 6.3 98.8 94.8 3.7 25.6 1.07 25 96.0 4.0 24.0 6.1 98.7 94.8 3.8 24.9 1.04

TABLE-US-00031 TABLE 31 Filter Element of Example 26 386 g/L Challenge Load Filtration Performance Final Final Ratio of Final Initial Initial Aggregated Monomeric Monomeric Aggregated Composition IgG Monomeric Aggregated Initial IgG IgG IgG IgG Final Ratio to Initial Solution IgG IgG Composition Removed Yield Recovered Recovered Composition Composition Number (%) (%) Ratio (%) (%) (%) (%) Ratio Ratio 1 97.1 2.9 33.5 90.8 89.6 87.0 0.3 290.0 8.66 2 97.1 2.9 33.5 43.1 96.1 93.3 1.7 54.9 1.64 3 96.6 3.4 28.4 15.9 98.1 94.8 2.9 32.7 1.15 6 96.8 3.2 30.3 95.1 81.8 79.2 0.2 396.0 13.07 7 96.5 3.5 27.6 21.3 98.0 94.6 2.8 33.8 1.22 8 98.4 1.6 61.5 2.6 98.4 96.8 1.6 60.5 0.98 18 96.0 4.0 24.0 59.1 91.8 88.1 1.6 55.1 2.30 19 96.2 3.8 25.3 8.3 99.1 95.3 3.5 27.2 1.08 20 96.2 3.8 25.3 4.4 99.3 95.5 3.6 26.5 1.05 23 96.2 3.8 25.3 34.7 92.5 89.0 2.5 35.6 1.41 24 96.0 4.0 24.0 4.7 98.8 94.8 3.8 24.9 1.04 25 96.0 4.0 24.0 2.4 99.3 95.3 3.9 24.4 1.02

TABLE-US-00032 TABLE 32 Filter Element of Example 27 386 g/L Challenge Load Filtration Performance Final Final Ratio of Final Initial Initial Aggregated Monomeric Monomeric Aggregated Composition IgG Monomeric Aggregated Initial IgG IgG IgG IgG Final Ratio to Initial Solution IgG IgG Composition Removed Yield Recovered Recovered Composition Composition Number (%) (%) Ratio (%) (%) (%) (%) Ratio Ratio 1 97.1 2.9 33.5 51.6 96.0 93.2 1.4 66.6 1.98 2 97.1 2.9 33.5 14.4 97.9 95.1 2.5 38.0 1.14 3 96.6 3.4 28.4 11.9 98.2 94.9 3.0 31.6 1.11 6 96.8 3.2 30.3 71.0 93.4 90.4 0.9 100.4 3.31 7 96.5 3.5 27.6 18.9 98.2 94.8 2.8 33.8 1.21 8 98.4 1.6 61.5 4.7 98.8 97.2 1.5 64.8 1.05 18 96.0 4.0 24.0 30.0 96.7 92.8 2.8 33.1 1.38 19 96.2 3.8 25.3 6.4 98.5 94.8 3.6 26.3 1.04 20 96.2 3.8 25.3 5.7 98.8 95.0 3.6 26.4 1.04 23 96.2 3.8 25.3 19.6 96.1 92.4 3.1 29.8 1.18 24 96.0 4.0 24.0 5.2 98.6 94.7 3.8 24.9 1.04 25 96.0 4.0 24.0 4.5 99.0 95.0 3.8 25.0 1.04

TABLE-US-00033 TABLE 33 Filter Element of Example 28 386 g/L Challenge Load Filtration Performance Final Final Ratio of Final Initial Initial Aggregated Monomeric Monomeric Aggregated Composition IgG Monomeric Aggregated Initial IgG IgG IgG IgG Final Ratio to Initial Solution IgG IgG Composition Removed Yield Recovered Recovered Composition Composition Number (%) (%) Ratio (%) (%) (%) (%) Ratio Ratio 1 97.1 2.9 33.5 73.7 92.1 89.4 0.8 111.7 3.33 2 97.1 2.9 33.5 16.0 98.1 95.3 2.4 39.7 1.19 3 96.6 3.4 28.4 10.0 98.5 95.2 3.1 30.7 1.08 6 96.8 3.2 30.3 83.0 84.7 82.0 0.5 164.0 5.41 7 96.5 3.5 27.6 16.1 98.7 95.2 2.9 32.8 1.19 8 98.4 1.6 61.5 1.1 99.2 97.6 1.6 61.0 0.99 18 96.0 4.0 24.0 35.4 94.0 90.2 2.6 34.7 1.45 19 96.2 3.8 25.3 3.3 99.3 95.5 3.7 25.8 1.02 20 96.2 3.8 25.3 2.2 99.9 96.1 3.7 26.0 1.03 23 96.2 3.8 25.3 27.5 92.8 89.3 2.8 31.9 1.26 24 96.0 4.0 24.0 1.6 99.7 95.7 3.9 24.5 1.02 25 96.0 4.0 24.0 1.7 100.0 96.0 3.9 24.6 1.02

TABLE-US-00034 TABLE 34 Filter Element of Example 29 387 g/L Challenge Load Filtration Performance Final Final Ratio of Final Initial Initial Aggregated Monomeric Monomeric Aggregated Composition IgG Monomeric Aggregated Initial IgG IgG IgG IgG Final Ratio to Initial Solution IgG IgG Composition Removed Yield Recovered Recovered Composition Composition Number (%) (%) Ratio (%) (%) (%) (%) Ratio Ratio 1 97.6 2.4 40.7 80.4 69.9 68.2 0.5 136.4 3.35 3 97.6 2.4 40.7 93.3 84.2 82.2 0.2 411.0 10.10 4 97.6 2.4 40.7 47.8 101.1 98.7 1.3 75.9 1.86 6 97.2 2.8 34.7 73.8 59.9 58.2 0.7 83.1 2.39 7 97.1 2.9 33.5 93.4 80.8 78.5 0.2 329.5 9.84 8 97.0 3.0 32.3 49.2 99.6 96.6 1.5 64.4 1.99 9 96.9 3.1 31.3 24.4 100.1 97.0 2.3 42.2 1.34 18 95.8 4.2 22.8 91.4 74.4 71.3 0.4 178.2 7.82 19 96.6 3.4 28.4 34.8 100.0 96.6 2.2 43.9 1.54 21 96.6 3.4 28.4 11.7 100.1 96.7 3.0 32.2 1.13 23 96.7 3.3 29.3 76.2 85.7 82.9 0.8 103.6 3.53 24 96.8 3.2 30.3 11.6 100.7 97.5 2.8 34.8 1.15 25 96.8 3.2 30.3 6.2 100.6 97.4 3.0 32.5 1.07

TABLE-US-00035 TABLE 35 Filter Element of Example 30 386 g/L Challenge Load Filtration Performance Final Final Ratio of Final Initial Initial Aggregated Monomeric Monomeric Aggregated Composition IgG Monomeric Aggregated Initial IgG IgG IgG IgG Final Ratio to Initial Solution IgG IgG Composition Removed Yield Recovered Recovered Composition Composition Number (%) (%) Ratio (%) (%) (%) (%) Ratio Ratio 1 96.6 3.4 28.4 68.3 73.1 70.6 1.1 64.2 2.26 2 96.8 3.2 30.3 83.8 78.2 75.7 0.5 151.4 5.00 3 96.7 3.3 29.3 80.7 90.0 87.0 0.6 145.0 4.95 4 96.8 3.2 30.3 11.9 98.3 95.2 2.8 34.0 1.12 6 96.8 3.2 30.3 73.3 70.9 68.6 0.9 76.2 2.51 7 96.4 3.6 26.8 76.2 91.5 88.2 0.9 98.0 3.65 8 95.1 4.9 19.4 6.3 97.2 92.4 4.6 20.1 1.04 9 97.1 2.9 33.5 3.2 99.1 96.2 2.8 34.3 1.02 11 95.8 4.2 22.8 92.5 80.6 77.2 0.3 257.3 11.29 12 95.1 4.9 19.4 5.7 97.4 92.6 4.6 20.1 1.04 13 96.4 3.6 26.8 2.2 99.2 95.6 3.5 27.3 1.02 14 96.2 3.8 25.3 0.9 99.4 95.6 3.8 25.1 0.99 16 95.2 4.8 19.8 74.8 87.8 83.6 1.2 69.7 3.52 17 95.1 4.9 19.4 1.9 99.4 94.5 4.8 19.7 1.01 18 95.1 4.9 19.4 1.3 99.3 94.4 4.8 19.7 1.01 19 96.0 4.0 24.0 1.8 99.7 95.7 3.9 24.5 1.02

[0331] The referenced descriptions contained in the patents, patent documents, and publications cited herein are incorporated by reference in their entirety as if each were individually incorporated. Various unforeseeable modifications and alterations to this disclosure will become apparent to those skilled in the art without departing from the scope and spirit of this disclosure. It should be understood that this disclosure is not intended to be unduly limited by the illustrative embodiments and examples set forth herein and that such examples and embodiments are presented by way of example only, with the scope of the disclosure intended to be limited only by the claims set forth herein as follows.