Email this article EFFECT OF METHODS OF IMMOBILIZATION OF ERYTHROPOIETIN ON THE SENSITIVITY FOR THE DETECTION OF SPECIFIC IgG TO EPO IN EXPERIMENTAL ANIMALS SERA
- Authors: Kudryashova A.M.1, Borisova O.V.1, Mikhailova N.A.1, Lonshakov D.V.2, Katlinsky A.V.2
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Affiliations:
- Mechnikov Research Institute of Vaccines and Sera
- Biopharmaceutical company LLC «Fort»
- Issue: Vol 94, No 6 (2017)
- Pages: 49-55
- Section: Articles
- URL: https://microbiol.crie.ru/jour/article/view/230
- DOI: https://doi.org/10.36233/0372-9311-2017-6-49-55
- ID: 230
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Abstract
Aim. The study of the effect of the erythropoietin coating procedure on sensitivity using the same secondary detection methods to quantify anti-EPO IgG positive animal sera. Materials and methods. Sera from experimental animals - rabbits and guinea pigs - after rhEPO injection were used. The methods includes directly coated ELISA and two types of immunochemical immobilization: capturing biotinylated rhEPO on streptavidin coated microtiier plates and capturing rhEPO via a specific antibody Results. Immunochemical rhEPO immobilization results in a sensitivity from 2 to 10 of magnitude higher than direct coating of rhEPO. Conclusion. Our findings show that the method of rhEPO immobilization to microtiter plates is a critical determinant for the sensitivity of ELISA used for measuring anti-EPO antibodies. Assays in which rhEPO was captured via a specific mAb, or in which biotinylated rhEPO was captured via streptavidin, are preferred to detect serum antibodies to native structural state.
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About the authors
A. M. Kudryashova
Mechnikov Research Institute of Vaccines and Sera
Author for correspondence.
Email: noemail@neicon.ru
Russian Federation
O. V. Borisova
Mechnikov Research Institute of Vaccines and Sera
Email: noemail@neicon.ru
Russian Federation
N. A. Mikhailova
Mechnikov Research Institute of Vaccines and Sera
Email: noemail@neicon.ru
Russian Federation
D. V. Lonshakov
Biopharmaceutical company LLC «Fort»
Email: noemail@neicon.ru
Russian Federation
A. V. Katlinsky
Biopharmaceutical company LLC «Fort»
Email: noemail@neicon.ru
Russian Federation
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