Construction of a combinatorial immune library of human single-chain antibodies to orthopoxviruses and selection of antibodies to recombinant prA30L of the variola virus

Viktoriya Dubrovskaya, A. B. Ulitin, A. G. Laman, I. P. Gileva, N. I. Bormotov, A. A. Ilyichev, F. A. Brovko, S. N. Shchelkunov, E. F. Belanov, N. V. Tikunova

Research output: Contribution to journalArticle

3 Scopus citations

Abstract

A combinatorial immune library of human single-chain antibodies (scAbs) was constructed using the genes coding for the variable domains of the heavy and light chains of human immunoglobulins. The genes were cloned from lymphocytes of four subjects vaccinated with the vaccinia virus (VACV). The library included 3 · 107 independent clones. After enrichment with clones producing scAbs against a recombinant analog of the variola virus envelope protein prA30L, the library was used to select a panel of scAbs binding both prA30L and VACV. All scAbs selected were tested for virus-neutralizing activity, and two scAbs proved to suppress VACV plaque formation in monolayers of Vero E6 cells. The specificity of antigen binding was verified by ELISA and Western blotting. The amino acid sequences of the virus-neutralizing scAbs were determined by sequencing their genes.

Original languageEnglish (US)
Pages (from-to)157-167
Number of pages11
JournalMolecular Biology
Volume41
Issue number1
DOIs
Publication statusPublished - Feb 1 2007
Externally publishedYes

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Keywords

  • Human single-chain antibodies
  • Phage display
  • Recombinant protein
  • Vaccinia virus
  • Variola virus
  • Virus neutralization

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology

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