EFFECT OF METHODS OF IMMOBILIZATION OF ERYTHROPOIETIN ON THE SENSITIVITY FOR THE DETECTION OF SPECIFIC IgG TO EPO IN EXPERIMENTAL ANIMALS SERA

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.