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Construction of immune library of murine immunoglobulin genes and screening of single-chain Fv-antibodies specific to diphtheria toxin B subunit

Oliĭnyk, O.S.; Kaberniuk, A.A.; Redchuk, T.A.; Korotkevych, N.V.; Labyntsev, A.Iu.; Romaniuk, S.I.; Kolybo, D.V.; Komisarenko, S.V.

Ukrains'kyi Biokhimichnyi Zhurnal 81(2): 68-79

2009

ISSN/ISBN: 0372-3909
PMID: 19873879
Document Number: 631805
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 10(7) to 10(9) M(-1).

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