Pavlova M.V., Gilchuk P.V., Pokholenko Ya.O., Nikolayev Yu.S., Kordium V.A.

Construction and characterization of immune cDNA combinatorial antibody library of mouse variable immunoglobuline genes

Cytology and Genetics 2008, v 42, number 2, 10-15

A cDNA combinatorial antibody library of mouse variable immunoglobulin fragments has been constructed from mice immunized with rhIFN-β1b. For this purpose, cDNAS of immunoglobulin variable heavy (V_H) and variable light (V_L) chains genes amplified from splenocytes were joined by linker DNA to form ScFv’s (single-chain Fv-antibodies). The obtained ScFv-DNA pool was cloned into a phagemid vector and used for Escherichia coli transformation. Using the phage display technique, bacterial clones producing single-chain antibodies specific to rhIFN-β1b were selected. The following characteristics of the combinatorial library were determined in this work: abundance, functional size, and the initial ScFv-DNA diversity in the library constructed. High specificity of interaction between phage displayed ScFv’s and rhIFN-β1b has been demonstrated.

Key words: cDNA combinatorial library, phage display, ScFv’s, e. coli, rhIFN-β1b