Recombinant polyclonal proteins targeting zika and methods of use thereof

Abstract

Provided herein are compositions comprising recombinant polyclonal protein (RPP) derived from mammalian plasma cells and plasmablasts. Specifically, RPPs specific to Zika virus are provided. Also provided are the methods of using the RPP for treatment of viral infection and the methods of making the RPPs.

1 . A pharmaceutical composition comprising:
 a mixture of at least 100 unique recombinant antibodies wherein the mixture is capable of neutralizing Zika virus with an IC50 lower than 0.001 mg/ml using an in vitro neutralization assay, and   a pharmaceutically acceptable excipient,   wherein the mixture of at least 100 unique recombinant antibodies has been generated by the process of
 a. isolating single cells from a blood sample from a donor exposed to an antigen of Zika virus; 
 b. amplifying polynucleotides, wherein each of the polynucleotides encodes a cognate pair of heavy chain and light chain variable regions from one of the single cells by overlap extension reverse transcriptase polymerase chain reaction (OE-RT-PCR); 
 c. cloning the polynucleotides obtained from the amplification into an expression vector, thereby obtaining constructs encoding antibody fragments; 
 d. expressing the antibody fragments from the constructs; 
 e. generating antibody expression constructs using the subset of the constructs, wherein each of the antibody expression constructs encodes a light chain variable region, a kappa or lambda-type light chain constant region, a heavy chain variable region, and a heavy chain constant region, 
 f. introducing the antibody expression constructs into a cell line, and 
 g. expressing antibodies from the antibody expression constructs in the cell line, thereby obtaining the at least 100 unique recombinant antibodies; 
   wherein each of the at least 100 unique recombinant antibodies is an antibody comprising a cognate pair of heavy chain and light chain variable regions from a single cell out of a blood sample from a donor exposed to an antigen of Zika virus.   
     
     
         2 . The pharmaceutical composition of  claim 1 , further comprising a compound selected from benzalkonium chloride, benzyl alcohol, benzoic acid, salicylic acid, thimerosal, phenethyl alcohol, methylparaben, propylparaben, chlorhexidine, sorbic acid and hydrogen peroxide in an amount effective to preserve the pharmaceutical composition. 
     
     
         3 . The pharmaceutical composition of  claim 1 , wherein each of the recombinant antibodies is a human IgG1 subtype. 
     
     
         4 . The pharmaceutical composition of  claim 1 , wherein each of the recombinant antibodies has been enriched based on neutralization activity against Zika virus. 
     
     
         5 . The pharmaceutical composition of  claim 1 , wherein the mixture comprises at least 1000 recombinant antibodies specifically binding to the antigen of Zika virus. 
     
     
         6 . The pharmaceutical composition of  claim 1 , wherein the mixture comprises at least 10,000 recombinant antibodies specifically binding to the antigen of Zika virus. 
     
     
         7 . The pharmaceutical composition of  claim 1 , wherein the antibody fragments are single-chain variable fragments (scFvs). 
     
     
         8 . The pharmaceutical composition of  claim 1 , formulated for intravenous infusion. 
     
     
         9 . The pharmaceutical composition of  claim 1 , formulated in a unit dose, wherein the unit dose is at least 0.1 g, 0.5 g, 1 g, 1.5 g, 2 g, 2.5 g, 3 g, 3.5 g, 4 g, 5 g, 10 g, 20 g, 30 g, 40 g, or 50 g of the at least 100 unique recombinant antibodies.