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ISSN 2410-776X (Online),
ISSN 2410-7751 (Print)

 3 2015

"Biotechnologia Acta" V. 8, No 3, 2015
https://doi.org/10.15407/biotech8.03.045
Р. 45-55, Bibliography 25, Ukrainian
Universal Decimal Classification: 557.161.2+612.017.1:616-097

SYNTHESIS OF 25-HYDROXYVITAMIN D3 CONJUGATE WITH KEYHOLE LIMPET HEMOCYANIN AND OBTAINING OF IMMUNE SERA

А. O. Mazanova, I. O. Shymanskyy, D. M. Petukhov, L. B. Drobot, M. M. Veliky, S. V. Komisarenko

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

The study was aimed at obtaining polyclonal antibodies that recognize 25-Hydroxyvitamin D3, for their further specifications and applications in immunochemical test systems of 25-Hydroxyvitamin D3, determination in blood serum. We used the methods of chemical synthesis of immunoconjugates (modified carbodiimide method using N´-ethyl carbodiimide), thin layer chromatography, gel filtration and indirect immunoenzyme analysis ELISA. The work describes the stages of the synthesis of 25-Hydroxyvitamin D3, immunoconjugate with keyhole limpet hemocyanin (KLH), which was used for receiving immune sera. As a result of mouse and rabbit immunization antisera were obtained and antibody titers against 25-Hydroxyvitamin D3, were tested by immunoenzyme assay. It was demonstrated that the titer of specific antibodies was higher in rabbits compared with mice. The resulting polyclonal antibodies can be used for the development of immunochemical test systems for screening studies of 25-Hydroxyvitamin D3, content in human blood serum as a marker of vitamin D3 availability.

Key words: 25-Hydroxyvitamin D3, haptens, immunoenzyme analysis, polyclonal antibodies.

© Palladin Institute of Biochemistry of the National Academy of Sciences of Ukraine, 2015