A METHOD FOR LINKING V-L AND V-H REGION GENES THAT ALLOWS BULK TRANSFER BETWEEN VECTORS FOR USE IN GENERATING POLYCLONAL IGG LIBRARIES

Libraries of Ab fragments have been produced by others from light and heavy chain cDNAs derived from populations of B lymphocytes and expres sed in bacteria. However, such libraries have not been transferred to eukaryotic expression vectors to generate polyclonal libraries of inta ct glycosylated Abs that can mediate effector functions. We present a method for transferring pairs of linked V-L-V-H region genes between c ircular prokaryotic and eukaryotic vectors. The key feature of the tra nsfer is that the V-L and V-H region genes are I in ked head to head ( <->) in opposite transcriptional orientations. To illustrate this meth od, a pair of V-L and V-H region cDNAs derived from an existing hybrid oma cell line were linked head to head by PCR, transferred as a unit b etween vectors, and expressed as an IgG Ab with Ag binding activity. A lthough we tested the transfer of a single V-L-V-H region gene pair, t his system is expected to allow the bulk transfer of physically linked V-L-V-H region gene combinations between different circular vectors a nd the expression of the same library as either Ab fragments or intact Abs.