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Home Archive 2012 № 6 ISOLATION AND CHARACTERISATION OF RECOMBINANT SINGLE CHAIN VARIABLE FRAGMENT ANTIBODIES (SCFV) AGAINST HUMAN HEPARINBINDING EGF-LIKE GROWTH FACTOR (HB-EGF) O. S. Oliinyk, A. A. Kaberniuk, D. V. Kolibo, S. V. Komisarenko
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ISSN 1995-5537

Scientific journal

БІОТЕХНОЛОГІЯ
V. 5, No. 6, 2012

«Біотехнологія» journal V. 5, No. 6, 2012
Р. 47-56, Bibliography 23, Ukrainian.
Universal Decimal classification: 602.68

ISOLATION AND CHARACTERISATION OF RECOMBINANT SINGLE CHAIN VARIABLE FRAGMENT ANTIBODIES (SCFV) AGAINST HUMAN HEPARINBINDING EGF-LIKE GROWTH FACTOR (HB-EGF)

O. S. Oliinyk, A. A. Kaberniuk, D. V. Kolibo, S. V. Komisarenko

Palladin Institute of Biochemistry of National Academy of Sciences of Ukraine, Kyiv

Heparin-binding epidermal growth factorlike growth factor (HB-EGF) is a member of the EGF family of growth factors, wich plays an important role in many physiological and pathological processes. Antibodies are an important tool for investigation of HB-EGF functions at normal and pathological processes. The aim of this study was to obtain and to study some properties of recombinant single-chain variable fragment (scFv) antibodies specific to HB-EGF. For this purpose a phage-displayed immune library of mice immunized with recombinant human soluble HB-EGF was generated. The library was screened against HB-EGF, and 2 groups of antibody clones with different fingerprinting patterns were isolated. Two clones — one from each group (scFv 5A and scFv 7E) were analyzed. It was shown that these scFv antibodies could bind both — human and mouse HB-EGF. Both clones recognized target antigen in Western blot, but only under non-reducing conditions. Specific binding of scFv 5A and scFv 7E to Vero cells with high level of HB-EGF on their surface was also demonstrated by confocal microscopy.

In this study scFv antibodies against soluble HB-EGF were obtained. They were suitable for ELISA, Western blot and immunocytochemistry applications. These scFv could be further used as a tool in HB-EGF research and, possibly, for the development of new diagnostic and therapeutic agents.

Key words: Heparin-binding epidermal growth factor-like growth factor, scFv-antibodies, phage display.

© Palladin Institute of Biochemistry of National Academy of Sciences of Ukraine, 2008

 

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Home Archive 2012 № 6 ISOLATION AND CHARACTERISATION OF RECOMBINANT SINGLE CHAIN VARIABLE FRAGMENT ANTIBODIES (SCFV) AGAINST HUMAN HEPARINBINDING EGF-LIKE GROWTH FACTOR (HB-EGF) O. S. Oliinyk, A. A. Kaberniuk, D. V. Kolibo, S. V. Komisarenko

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