OBTAINING OF RECOMBINANT HUMAN HEPARIN BINDING EGF-LIKE GROWTH FACTOR AND PERSPECTIVES OF ITS APPLICATION IN BIOTECHNOLOGY N. V. Korotkevich, D. V. Kolibo, А. J. Labyntsev, S. І. Romaniuk, S. V. Komisarenko

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ISSN 1995-5537

"Biotechnology" journal Vol. 3, No. 4, 2010
Р. 44-54, Bibliography 31, Ukranian
Universal Decimal classification: 577(112+354.9)+57(085.2+086.164)+ 616-006.6

OBTAINING OF RECOMBINANT HUMAN HEPARIN BINDING EGF-LIKE GROWTH FACTOR AND PERSPECTIVES OF ITS APPLICATION IN BIOTECHNOLOGY

N. V. Korotkevich, D. V. Kolibo, А. J. Labyntsev, S. І. Romaniuk, S. V. Komisarenko

Palladian Institute of Biochemistry of Ukrainian of National Academy of Sciences, Kyiv

In order to study functional activity and possibility of using in test-systems for diphtheria toxin (DT) detection the recombinant soluble form of human heparin binding EGF-like growth factor (sHB-EGF) was obtained. It was shown that recombinant sHB-EGF could bind DT and heparin and also stimulate 3Т3 mouse fibroblast’s proliferation in vitro. Affinity constant for the interaction of recombinant sHB-EGF with DT estimated by Friquet method was 1.67•10⁸ М^–1 that was similar to the affinity of natural form of sHB-EGF. Recombinant sHB-EGF was used in the immunoenzyme «sandwich»-analyses instead capture antibodies for DT detection. Sensitivity of the developed method was 1.9 ng/ml. Obtained potential biotechnological product probably can be used as component of the test-systems for diphtheria diagnosis, as component of therapeutic drugs for stimulation of tissue regeneration or as a tool for fundamental researches.

Key words: human heparin binding EGF-like growth factor, soluble form, recombinant protein, test-system, diphtheria toxin.