Open AccessEFFECT OF METHODS OF IMMOBILIZATION OF ERYTHROPOIETIN ON THE SENSITIVITY FOR THE DETECTION OF SPECIFIC IgG TO EPO IN EXPERIMENTAL ANIMALS SERA
Author(s) -
A. M. Kudryashova,
О. В. Борисова,
Н. А. Михайлова,
Dmitry V. Lonshakov,
A. V. Katlinsky
Publication year - 2017
Publication title -
žurnal mikrobiologii, èpidemiologii i immunobiologii
Language(s) - English
Resource type - Journals
eISSN - 2686-7613
pISSN - 0372-9311
DOI - 10.36233/0372-9311-2017-6-49-55
Subject(s) - biotinylation , erythropoietin , streptavidin , antibody , chemistry , chromatography , microbiology and biotechnology , biotin , immunology , medicine , biology , biochemistry
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.