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Construction and characterization of immune cDNA combinatorial antibody library of mouse variable immunoglobuline genes
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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 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
E-mail: mariaka ukr.net, gilchuk ukr.net
|Coded & Designed by Volodymyr Duplij||Modified 18.09.21|