TSitologiya i Genetika 2008, vol. 42, no. 2, 10-15
Cytology and Genetics , vol. , no. , , doi: https://www.doi.org/

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

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

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 (VH) and variable light (VL) 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.

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

TSitologiya i Genetika
2008, vol. 42, no. 2, 10-15

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