| CPC C07K 16/1081 (2013.01) [A61P 31/14 (2018.01); C07K 2317/52 (2013.01); C07K 2317/565 (2013.01); C07K 2317/622 (2013.01)] | 6 Claims |
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1. A method of treating a subject in need thereof, the method comprising administering to the subject an effective amount of 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.
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