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Characterization of a panel of mouse single-chain antibodies against recombinant human interferon β1b<
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A panel of single-chain antibodies (ScFv’s – single-chain Fv-antibodies) against recombinant human interferon beta 1b (rhIFN-β1b) has been obtained from immune and naïve combinatorial cDNA libraries of the mouse variable immunoglobulin genes. ScFv’s were expressed into Escherichia coli cells. For producers isolated from the immune library a difference in production yield of ScFv’s in periplasm and incubation medium as well as their expression stability in passages and storage stability have been demonstrated. After sequencing of target DNA the multiple alignment and structural analysis of ScFv’s sequences with different primary structures were carried out and significant difference in both complementarity-determining (CDR) and framework (FR) regions of their variable domains has been shown. For the ScFv’s isolated from the immune library, specificity of their binding with native and denatured rhIFN-β1b in ELISA and Western-blotting as well as their high storage stability have been shown. The affinity constants for each representatives of the ScFv’s panel were in the range from 1.9610-8 to 1.6910-9 M.
Key words: cDNA library, ScFv’s, Escherichia coli, rhIFN-β1b, specificity, affinity
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|Coded & Designed by Volodymyr Duplij||Modified 06.12.23|