Construction of combinatorial immune library of single chain human antibodies to orthopoxviruses and selection from this library antibodies to recombinant protein prA30L of variola virus

V. V. Dubrovskaia, A. B. Ulitin, A. G. Laman, I. P. Gileva, N. I. Bormotov, A. A. Il'ichev, F. A. Brovko, S. N. Shchelkunov, E. F. Belanov, N. V. Tikunova

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3 Scopus citations

Abstract

A combinatorial immune library of human single-chain antibody fragments (scFv) was constructed on the base of genes encoding variable domains of heavy and light chains of immunoglobulins cloned from the lymphocytes of four vaccinia virus (VACV) vaccinated donors. The size of the library was 3 x 10(7) independent clones. After the library was enriched with the clones producing scFv against recombinant analogue of variola virus surface protein prA30L, a panel of unique antibodies specific to both prA30L and VACV was selected from the library. A plaque reduction neutralization test was performed for all selected antibodies and two antibodies were shown to be able to neutralize plaque formation of VACV in Vero E6 cells monolayer. Binding specificities of these antibodies were confirmed using ELISA and Western blot analysis. To determine the amino acid sequences of neutralizing antibodies their genes were sequenced.

Original languageEnglish (US)
Pages (from-to)173-185
Number of pages13
JournalMolekuliarnaia biologiia
Volume41
Issue number1
StatePublished - Jan 1 2007

ASJC Scopus subject areas

  • Medicine(all)

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    Dubrovskaia, V. V., Ulitin, A. B., Laman, A. G., Gileva, I. P., Bormotov, N. I., Il'ichev, A. A., Brovko, F. A., Shchelkunov, S. N., Belanov, E. F., & Tikunova, N. V. (2007). Construction of combinatorial immune library of single chain human antibodies to orthopoxviruses and selection from this library antibodies to recombinant protein prA30L of variola virus. Molekuliarnaia biologiia, 41(1), 173-185.