The present invention provides engineered DNase proteins (including DNase1-like 3 and DNase1) that are useful for treating conditions characterized by neutrophil extracellular trap (NET) accumulation and/or release. In some aspects, the invention provides compositions and methods for preventing or treating vascular occlusion involving NETs. As demonstrated herein, NETs participate in a non-canonical mechanism for vascular occlusion, which is not dependent on fibrin or platelets.

The present invention includes compositions and methods for the expression, secretion and use of novel compositions for use as, e.g., vaccines and antigen delivery vectors, to delivery antigens to antigen presenting cells. In one embodiment, the vector is an anti-CD40 antibody, or fragments thereof, and one or more antigenic peptides linked to the anti-CD40 antibody or fragments thereof, including humanized antibodies.

Provided are fusion proteins including an amino acid sequence of an ectodomain of Spike protein of a coronavirus, such as SARS-CoV-2, joined to an amino acid sequence of a ferritin subunit polypeptide. Nanoparticles including such fusion proteins, with surface-exposed trimers of the ectodomain of the Spike protein of the coronavirus, are also provided. Also provided are nucleic acids and vectors encoding the fusion proteins, cells containing such nucleic acid and vectors, immunogenic compositions including the fusion proteins, the nanoparticles, or the vectors, as well as corresponding methods and kits.

The disclosure provides materials in the form of flavivirus variants that each encode a Non-Structural Protein-1 (NS1) variant, wherein the coding region is a chimera of at least two different NS1 coding regions, or wherein the coding region has at least one mutation in a codon of a canonical Asn-Xxx-Ser/Thr N-linked glycosylation site, wherein Asn is asparagine, Xxx is any amino acid, and Ser/Thr is either serine or threonine, or wherein the coding region is both a chimera and has at least one mutation in a codon of a canonical N-liked glycosylation site, wherein Asn is asparagine, Xxx is any amino acid, and Ser/Thr is either serine or threonine. The disclosure also provides methods of using such flavivirus variants to inhibit the transmission of infectious flavivirus.

The present invention includes an immunogenic protein, nucleic acid, plant and immunization comprising a fusion protein that has at least 90% amino acid identity to an amino acid sequence of a modified thermostable lichenase (LicKM) polypeptide as set forth in SEQ ID NO:9, wherein the LicKM polypeptide comprises an N-terminus, a C-terminus, and an inner loop region, and wherein a Receptor Binding Domain (RBD) or a Receptor Binding Motif (RBM) of a coronavirus spike protein is positioned at, at least one of, the N-terminus, the C-terminus, or in a loop region of the LicKM polypeptide.

Methods for glycosylation of Grp94 chaperone proteins, including Grp94 fragments, mutants, and fusion proteins, expressed in mammalian cells are presented. The method comprises altering a Pre-N domain of the Grp94 chaperone protein, wherein the alteration of the Pre-N domain of the Grp94 chaperone protein leads to glycosylation at available glycosylation sites downstream of the Pre-N domain of the Grp94 chaperone protein, when expressed in mammalian cells.

The present invention includes compositions and methods for the expression, secretion and use of novel compositions for use as, e.g., vaccines and antigen delivery vectors, to delivery antigens to antigen presenting cells. In one embodiment, the vector is an anti-CD40 antibody, or fragments thereof, and one or more antigenic peptides linked to the anti-CD40 antibody or fragments thereof, including humanized antibodies.

The present invention includes compositions and methods for the expression, secretion and use of novel compositions for use as, e.g., vaccines and antigen delivery vectors, to delivery antigens to antigen presenting cells. In one embodiment, the vector is an anti-CD40 antibody, or fragments thereof, and one or more antigenic peptides linked to the anti-CD40 antibody or fragments thereof, including humanized antibodies.

Disclosed are methods, systems, components, and compositions for cell-free synthesis of glycosylated proteins. The glycosylated proteins may be utilized in vaccines, including anti-bacterial vaccines. The glycosylated proteins may include a bacterial polysaccharide conjugated to a carrier, which may be utilized to generate an immune response in an immunized host against the polysaccharide conjugated to the carrier. The glycosylated proteins may be synthesized in cell-free glycoprotein synthesis (CFGpS) systems using prokaryote cell lysates that are enriched in components for glycoprotein synthesis such as oligosaccharyltransferases (OSTs) and lipid-linked oligosaccharides (LLOs) including OSTs and LLOs associated with synthesis of bacterial O antigens.

Provided is a technique for highly expressing a target protein in a plant cell by using a glycosylation domain, a recombinant vector comprising a gene encoding a fusion protein of a glycosylation domain and a target protein, a recombinant cell, a transformed plant, and a method of producing a target protein using these.