Disclosed herein is an activatable fusion protein that includes, in an N- to C-terminal direction, an albumin, a matrix metalloproteinase-cleavable linker, and an immunoadhesin which includes a cytotoxic T lymphocyte-associated antigen-4 (CTLA4) having an N-terminal extracellular domain and an IgG Fc region, wherein the albumin is released from the activatable fusion protein in the presence of a matrix metalloproteinase that cleaves the cleavable linker, so that the N-terminal extracellular domain of the CTLA4 binds to CD80 or CD86. Also disclosed herein is use of the activatable fusion protein for suppressing a T cell-dependent immune response.

Provided are methods and compositions comprising ulinastatin polypeptides for treating diseases associated with elevated neutrophil elastase (NE), including NE-associated lung and skin diseases.

Provided are methods and compositions comprising ulinastatin polypeptides for treating diseases associated with elevated neutrophil elastase (NE), including NE-associated lung and skin diseases.

The present disclosure provides a library of engineered DNASE proteins (including DNASE1, DNASE1-LIKE 1, DNASE1-LIKE 2, DNASE1-LIKE 3, DNASE2A, DNASE2B) that allows to select drug candidates for developing therapeutics for treating conditions characterized by neutrophil extracellular trap (NET) accumulation and/or release. In accordance with the invention, the selected DNase variant has improved properties, including properties amenable to clinical development, including manufacturing, toxicology, pharmacokinetic, and/or use in therapy.

The present disclosure provides a library of engineered DNASE proteins (including DNASE1, DNASE1-LIKE 1, DNASE1-LIKE 2, DNASE1-LIKE 3, DNASE2A, DNASE2B) that allows to select drug candidates for developing therapeutics for treating conditions characterized by neutrophil extracellular trap (NET) accumulation and/or release. In accordance with the invention, the selected DNase variant has improved properties, including properties amenable to clinical development, including manufacturing, toxicology, pharmacokinetic, and/or use in therapy.

Compositions that include an albumin binding domain and a fusion partner (e.g., a cytokine or a binding moiety) are provided. Such therapeutics have increased serum half-life and find use in applications where one or more such therapeutics are needed, for example, in oncology applications.

A method for producing a Wnt protein-afamin complex, the method comprising the steps of culturing Wnt protein-expressing cells in a culture medium containing a purified afamin or a recombinant afamin or co-culturing Wnt protein-expressing cells and recombinant afamin-expressing cells or culturing cells expressing both a Wnt protein and afamin; obtaining the culture supernatant; and optionally performing affinity purification to obtain the Wnt protein-afamin complex from the culture supernatant.

A method for producing a Wnt protein-afamin complex, the method comprising the steps of culturing Wnt protein-expressing cells in a culture medium containing a purified afamin or a recombinant afamin or co-culturing Wnt protein-expressing cells and recombinant afamin-expressing cells or culturing cells expressing both a Wnt protein and afamin; obtaining the culture supernatant; and optionally performing affinity purification to obtain the Wnt protein-afamin complex from the culture supernatant.

A method of preparing a conditionally active polypeptide from a parent polypeptide, comprising steps of evolving a DNA encoding the parent polypeptide by increasing a net charge of the parent polypeptide using one or more techniques selected from increasing a total number of codons of charged amino acid residues in the DNA and decreasing a total number of codons of uncharged amino acid residues in the DNA to create mutant DNAs; expressing the mutant DNAs to obtain mutant polypeptides; and selecting the conditionally active polypeptide from the mutant polypeptides which exhibits a decrease in activity in a first assay at a first value of a condition compared to the same activity in a second assay at a second value of the same condition. The conditionally active polypeptide, pharmaceutical compositions containing same, nanoparticle and drug conjugates thereof and uses thereof are also provided.

A method of preparing a conditionally active polypeptide from a parent polypeptide, comprising steps of evolving a DNA encoding the parent polypeptide by increasing a net charge of the parent polypeptide using one or more techniques selected from increasing a total number of codons of charged amino acid residues in the DNA and decreasing a total number of codons of uncharged amino acid residues in the DNA to create mutant DNAs; expressing the mutant DNAs to obtain mutant polypeptides; and selecting the conditionally active polypeptide from the mutant polypeptides which exhibits a decrease in activity in a first assay at a first value of a condition compared to the same activity in a second assay at a second value of the same condition. The conditionally active polypeptide, pharmaceutical compositions containing same, nanoparticle and drug conjugates thereof and uses thereof are also provided.