Ukr.Biochem.J. 2015; Volume 87, #2, March-April

Recombinant single chain variable fragment antibodies (scFv) against Pro144-Leu155 fragment of human protein C

O. S. Oliinyk, K. O. PalyvodA, N. E. Lugovskaya,
D. V. Kolibo, E. V. Lugovskoy, S. V. Komisarenko

Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: lenaoliinyk@mail.ru

The aim of this work was to obtain the recombinant single chain variable fragments of antibodies (scFv) against human protein C, the key component of blood anticoagulation system. For this purpose a peptide that mimics a Pro144-Leu155 sequence of protein C was synthesized and the murine immune scFv library against this peptide was constructed. The protein C specific scFv 9E were selected from the constructed library by the phage-display method. The scFv 9E dissociation constant was found to be 2∙10-9 М. It was shown that scFv 9E were suitable for protein C detection by ELISA and Western blotting. Selected scFv could be further used for protein C investigation and for the development of quantitative methods for protein C detection in human blood.

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