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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.

 


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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 ScFvs (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 ScFvs and rhIFN-β1b has been demonstrated.

Key words: cDNA combinatorial library, phage display, ScFvs, e. coli, rhIFN-β1b

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

E-mail: mariaka ukr.net, gilchuk ukr.net

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, Tsitol Genet., 2008, vol. 42, no. 2, pp. 10-15.




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