Open AccessEnzyme immunoassay of vitamin d3 metabolites
Author(s) -
А. C. Круглик,
Dmitrii V. Tsavlovskii,
Т. В. Миранцова,
О. Л. Шарко,
Vadim V. Shmanai
Publication year - 2020
Publication title -
vescì nacyânalʹnaj akadèmìì navuk belarusì. seryâ hìmìčnyh navuk
Language(s) - English
Resource type - Journals
eISSN - 2524-2342
pISSN - 1561-8331
DOI - 10.29235/1561-8331-2020-56-2-197-205
Subject(s) - hapten , polyclonal antibodies , horseradish peroxidase , bovine serum albumin , immunoassay , conjugate , metabolite , chemistry , vitamin , enzyme , cholecalciferol , chromatography , vitamin d and neurology , active metabolite , peroxidase , vitamin d binding protein , serum albumin , biochemistry , antibody , biology , immunology , endocrinology , mathematical analysis , mathematics
We report herein improved version of the synthesis of hapten based on cholecalciferol and its active metabolite 25-hydroxycholecalcalferol. The methodology of obtaining high-molecular immunogenic conjugates of vitamin D 3 derivatives with bovine serum albumin and horseradish peroxidase conjugates for direct ELISA was optimized. Immunisation of rabbits was carried out and polyclonal antibodies to 25-hydroxycholecalceferol were obtained and tested in an enzyme-linked immunosorbent assay. To improve the accuracy of the method, the sample preparation procedure was optimized, including the release of vitamin D 3 and its active metabolites from complexes with vitamin D-binding protein.