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"10.29235/1561-8331-2018-54-1-72-79vestich-305Research ArticleБИООРГАНИЧЕСКАЯ ХИМИЯBIOORGANIC CHEMISTRYПОЛУЧЕНИЕ БЕЛКОВЫХ КОНЪЮГАТОВ И МОНОКЛОНАЛЬНЫХ АНТИТЕЛ ДЛЯ ИММУНОФЕРМЕНТНОГО АНАЛИЗА ТЕТРАЦИКЛИНАPREPARATION OF PROTEIN CONJUGATES AND MONOCLONAL ANTIBODIES FOR IMMUNOASSAY OF TETRACYCLINEКругликА. С.KruhlikA. S.

науч. сотрудник

ул. Сурганова, 13, 220072, Минск

Researcher

13, Surganov Str., 220072, Minsk

a.kruhlik@gmail.comАкаловичС. Т.AkalovichS. T.

канд. биол. наук, ст. науч. сотрудник

Долгиновский тракт, 160, 220053, Минск

Ph. D. (Biology), Senior researcher, Research and Production Center for Hematology and Medical Biotechnology

160, Dolginovski tract, 220053, Minsk

svkoleda@mail.ruШаркоО. Л.SharkoO. L.

канд. хим. наук, ст. науч. сотрудник

ул. Сурганова, 13, 220072, Минск

Ph. D. (Chemistry), Senior researcher

13, Surganov Str., 220072, Minsk

sharko@ifoch.bas-net.byШманайВ. В.ShmanaiV. V.

канд. хим. наук, зав. лаб. химии биоконъюгатов

ул. Сурганова, 13, 220072, Минск

Ph. D. (Chemistry), Head of the Laboratory of Bioconjugate Chemistry

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 BelarusРеспубликанский научно-практический центр трансфузиологии и медицинских биотехнологийInstitute of Physical Organic Chemistry of the National Academy of Sciences of BelarusИнститут физико-органической химии Национальной академии наук БеларусиThe Republic Research & Production Center for Transfusiology and Medical Biotechnologies2018050320185417279Copyright © Круглик А.С., Акалович С.Т., Шарко О.Л., Шманай В.В., 20182018Круглик А.С., Акалович С.Т., Шарко О.Л., Шманай В.В.Kruhlik A.S., Akalovich S.T., 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/305

Разработаны методы контролируемой химической модификации бычьего сывороточного альбумина и пероксидазы хрена, позволяющие вводить в белок как минимальное число модификаций для сохранения биологической активности в иммуноанализе, так и максимальное – для получения иммуногенов, а также варьировать линкер и сайт связывания антигена с белком для повышения специфичности иммунного ответа. Получены и охарактеризованы белковые конъюгаты тетрациклина. Проведена иммунизация мышей, выделены моноклональные антитела к тетрациклину, которые испытаны в иммуноферментном анализе тетрациклина. Чувствительность определения составила IC50=5 мкг/мл. 

We report herein on the methods to introduce a desirable number of modifications into bovine serum albumin and horseradish peroxidase in a controlled manner. The low modification to protein ratio is required when the biological activity of the enzyme has to be preserved, and the high modification to protein ratio is needed when producing immunogens. By varying the linker and the site of antigen binding with the carrier protein, we were able to produce the antibodies of high specificity. Protein conjugates of tetracycline were synthesized and characterized. Immunization of mice was carried out and monoclonal antibodies for immunoassay of tetracycline were obtained and tested in ELISA, IC50=5 ug/ml. 

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