A method for producing a recombinant allotype-specific rabbit monoclonal antibody

摘要

A method for producing a full length coding sequence for a b4 allotype rabbit monoclonal antibody is provided. In some embodiments, this method may involve: providing a rabbit antibody-producing B-lymphocyte having a b4 allotype; fusing the B-lymphocyte with a rabbit fusion partner or derivative thereof having an allotype other than b4 (e.g. 240E cells) to produce a hybridoma; isolating cDNA from the hybridoma; amplifying full length heavy and light chain domains from the cDNA using (i) a light chain primer pair that comprises a forward primer having a 3 end of sequence according to SEQ ID No: 1 and a reverse primer having a 3 end of sequence according to SEQ ID No: 2, and (ii) a heavy chain primer pair that comprises a forward primer having a 3 end of sequence according to SEQ ID No: 3 and a reverse primer having a 3 end of sequence according to SEQ ID No: 4; cloning the amplified sequence into an expression vector to produce a first plasmid transfecting the plasmids into a mammalian host cell; culturing and purifying the antibodies produced. Related isolated primers, compositions, and uses of the primers are also claimed.