eISSN: 1644-4124
ISSN: 1426-3912
Central European Journal of Immunology
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2/2017
 
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abstract:
Experimental immunology

New human single chain anti-idiotypic antibody against benzo[a]pyrene

Artem E. Studennikov, Valentin A. Ustinov, Vera V. Morozova, Nina V. Tikunova, Andrey N. Glushkov

(Cent Eur J Immunol 2017; 42 (2): 123-130)
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The naïve library from the lymphocytes of healthy humans was screened by murine single-stranded idiotypic antibodies against benzo[a]pyrene (pSh). The phage clone which contained of anti-idiotypic antibody against benzo[a]pyrene, designated as A4, was chosen for further work because of highly specific to pSh. The available protein databases were searched. The A4 amino acid sequence was unique and 76% identical to a sequence in antibody against interferon g. The A4 protein was expressed in bacteria and purified by two different methods: His-tagged A4 and CBD-fusion A4. Both the A4 bound to pSh and also to the human single chain idiotypic antibody against the benzo[a]pyrene (T72) by ELISA. The Kd values of A4 for pSh and T72 were very close: 4.44 × 10-7 M and 5.71 × 10-7M, respectively. A4 was a competitor with benzo[a]pyrene for binding sites of both idiotypic pSh and T72 in competitive ELISA. Thus, A4 was a high affinity anti-idiotypic against benzo[a]pyrene which recognised pSh and T72 active sites.
keywords:

anti-idiotypic antibody, benzo[a]pyrene, immunology, phage display, polycyclic aromatic hydrocarbons, single-chain antibody

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