Journal archive > 2009 > N 2 March-April

CONSTRUCTION OF IMMUNE LIBRARY OF MURINE IMMUNOGLOBULIN GENES
AND SCREENING OF SINGLE-CHAIN Fv-ANTIBODIES TO DIPHTHERIA TOXIN B SUBUNIT
 
O. S. Oliinyk, A. A. Kaberniuk, T.?A.?Redchuk, N. V. Korotkevich, A.?J.?Labyntsev,

S. I. Romanyuk, D.?V.?Kolibo, S. V. Komisarenko

Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: This email address is being protected from spambots. You need JavaScript enabled to view it.

The aim of this work was to obtain the panel of recombinant single-chain Fv-antibodies against diphtheria toxin B subunit, the main diagnostic and pathogenic antigen of Corynebacterium diphtheriae. For this purpose we have constructed the immune library of murine immunoglobulin genes. A number of scFv specific to diphtheria toxin B subunit were acquired from the obtained library after one round of selection by phage-display. ScFv encoding DNA-fragments of eight clones were subcloned into plasmid pET-22b(+). It was shown that selected scFv were highly specific to diphtheria toxin B subunit, with affinity constant for different clones ranged from 107 to 109 М-1.

Key words: scFv, single chain antibodies, library of immunoglobulin genes, phage display, diphtheria toxin.

The original article in Ukrainian is available for download in PDF format.

© The Ukrainian Biochemical Journal