vestichИзвестия Национальной академии наук Беларуси. Серия химических наукProceedings of the National Academy of Sciences of Belarus, Chemical Series1561-83312524-2342The Republican Unitary Enterprise Publishing House "Belaruskaya Navuka"10.29235/1561-8331-2020-56-2-197-205vestich-584Research ArticleБИООРГАНИЧЕСКАЯ ХИМИЯBIOORGANIC CHEMISTRYИммуноферментный анализ метаболитов витамина D3Enzyme immunoassay of vitamin d3 metabolitesКругликА. C.KruhlikA. S.

Круглик Александр Сергеевич – науч. сотрудник

ул. Сурганова, 13, 220072, Минск

Aliaksandr S. Kruhlik – Researcher

13, Surganov Str., 220072, Minsk

a.kruhlik@gmail.comЦавловскийД. В.TsaulouskiD. V.

Цавловский Дмитрий Валентинович – науч. сотрудник

ул. Сурганова, 13, 220072, Минск

Dzmitry V. Tsaulouski – Researcher

13, Surganov Str., 220072, Minsk

tsaulouski@gmail.comМиранцоваТ. В.MirantsovaT. V.

Миранцова Татьяна Валентиновна – сотрудник

Tatsiana V. Mirantsova – Researcher

hemma-test@yandex.ruШаркоО. Л.SharkoO. L.

Шарко Ольга Леонидовна – канд. хим. наук, вед. науч. сотрудник

ул. Сурганова, 13, 220072, Минск

Olga L. Sharko – Ph. D. (Chemistry), Leading Researcher

13, Surganov Str., 220072, Minsk

olsharko@gmail.comШманайВ. В.ShmanaiV. V.

Шманай Вадим Владимирович – канд. хим. наук, зав. лаб. 

ул. Сурганова, 13, 220072, Минск

Vadim V. Shmanai – Ph. D. (Chemistry), Head of the Laboratory

13, Surganov Str., 220072, Minsk

shmanai@ifoch.bas-net.byИнститут физико-органической химии Национальной академии наук БеларусиInstitute of Physical Organic Chemistry of the National Academy of Sciences of BelarusСООО Хемма-тестHemma-test Joint LLC202007062020562197205Copyright © Круглик А.C., Цавловский Д.В., Миранцова Т.В., Шарко О.Л., Шманай В.В., 20202020Круглик А.C., Цавловский Д.В., Миранцова Т.В., Шарко О.Л., Шманай В.В.Kruhlik A.S., Tsaulouski D.V., Mirantsova T.V., Sharko O.L., Shmanai V.V.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://vestichem.belnauka.by/jour/article/view/584

Описан улучшенный вариант синтеза гаптена на основе холекальциферола и его активного метаболита 25-гидроксихолекальцеферола. Оптимизирована методика получения высокомолекулярных иммуногенных конъюгатов производных витамина D3 с бычьим сывороточным альбумином, а также конъюгатов с пероксидазой хрена для прямого ИФА. Проведена иммунизация кроликов, выделены поликлональные антитела к 25-гидроксихолекальцеферолу, которые испытаны в иммуноферментном анализе. Для повышения точности метода оптимизирован процесс пробоподготовки, включающий высвобождение витамина D3 и его активных метаболитов из комплексов с витамин D-связывающим белком.

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 D3 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 D3 and its active metabolites from complexes with vitamin D-binding protein.

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The authors declare that there are no conflicts of interest present.