Generation and characterization of an immune combinatorial cDNA library of variable genes of mouse immunoglobulins |
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| Authors: | M V Pavlova P V Gilchuk Ia O Pokholenko I S Nikolaiev and V A Kordium |
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| Institution: | (1) Taras Shevchenko National University of Kyiv, Kyiv, Ukraine;(2) Institute of Molecular Biology and Genetics, National Academy of Sciences of Ukraine, Kyiv, Ukraine |
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| Abstract: | A combinatorial cDNA library of mouse variable immunoglobulin genes from mice immunized with recombinant human interferon
β1b (rhIFN-β1b) has been constructed. For this purpose, cDNA of variable genes of heavy (V
H
) and light (V
L
) immunoglobulin chains amplified from splenocytes were joined by linker DNA to form single-chain antibodies (single-chain
Fv-antibodies, or ScFv’s). The ScFv-DNA pool thus obtained was cloned into a phagemid vector and used to transform Escherichia coli. Bacterial clones that produce single-chain antibodies that are specific to rhIFN-β1b ScFv’s were selected using the technique
of phage display. Such characteristics of generated library as abundance, functional size, and initial diversity of the ScFv-DNA
sequences were established in the study. A high degree of specificity of interaction of selected phage displayed ScFv’s with
rhIFN-β1b has been demonstrated.
Original Ukrainian Text ? M.V. Pavlova, P.V. Gilchuk, Ia. O. Pokholenko, I.S. Nikolaiev, V.A. Kordium, 2008, published in
Tsitologiya i Genetika, 2008, Vol. 42, No. 2, pp. 10–15. |
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