Patent ID: 8283294
Filing Date: 2012-10-09
Classification: C07K,C12N

Abstract:
1. A method of producing a library of cognate pairs comprising linked immunoglobulin variable region encoding sequences, said method comprising: (a) immunizing a rodent with a target antigen and providing a lymphocyte-comprising cell fraction from said rodent; (b) isolating from said lymphocyte-comprising cell fraction cells expressing CD43 and CD138 antigen or MHCII and B220 antigen; (c) distributing the isolated cells from step b) individually into a plurality of vessels to obtain a population of isolated single cells, wherein each cell expresses CD43 and CD138 antigen or MHCII and B220 antigen; and (d) amplifying, in a multiplex molecular amplification procedure, nucleotide sequences of interest using templates derived from said isolated single cells; and effecting linkage of the amplified nucleotide sequences of interest to generate a library of cognate pairs of a light chain variable region encoding sequence associated with a heavy chain variable region encoding sequence.