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repertoire cloning

A method to produce human monoclonal antibodies. mRNAs are isolated from a tissue that generates immunoglobulins (e.g. spleen) and a reverse transcriptase makes cDNA copies of them. The Fd regions, i.e. the H-chain component of the FAB fragment (the VH and CH1 domains of the immunoglobulins regions), are selected and amplified by PCR to obtain a heavy-chain library. Similarly, a light-chain library is obtained. The two populations of cDNA are combined into a random pairing to form combinatorial constructs, inserted into coliphage vector along with a promoter, and used to infect E. coli. Plaques are then screened for ability to bind an antigen. (see also synthetic peptide combinatorial library (SPCL))Barton, D.R. (1991) Trends Biotechnol. 9, 169-175


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