pharmjournalРазработка и регистрация лекарственных средствDrug development & registration2305-20662658-5049LLC «CPHA»10.33380/2305-2066-2024-13-2-1507pharmjournal-1808Research ArticleДОКЛИНИЧЕСКИЕ И КЛИНИЧЕСКИЕ ИССЛЕДОВАНИЯPRECLINICAL AND CLINICAL STUDIESВалидация тест-системы на основе метода иммуноферментного анализа для определения концентрации бевацизумаба в биологических жидкостяхValidation of an enzyme-linked immunosorbent assay test-system for bevacizumab concentration determing in biological fluidshttps://orcid.org/0000-0003-3212-4369ПисаревВ. В.PisarevV. V.

111024, г. Москва, ул. 5-я Кабельная, д. 2-Б, стр. 1, офис 3–1

office 3–1, building 1, 2-B, 5th Kabelnaya str., Moscow, 111024

vladimir.pisarev@probiotech.ruhttps://orcid.org/0000-0002-1676-7754ИвановА. В.IvanovA. V.

111024, г. Москва, ул. 5-я Кабельная, д. 2-Б, стр. 1, офис 3–1

office 3–1, building 1, 2-B, 5th Kabelnaya str., Moscow, 111024

gostyatin@gmail.comОбщество с ограниченной ответственностью «Научно-производственный центр Пробиотек»LLC "Probiotech"202426042024132181189Copyright © Писарев В.В., Иванов А.В., 20242024Писарев В.В., Иванов А.В.Pisarev V.V., Ivanov A.V.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.pharmjournal.ru/jour/article/view/1808Введение

Введение. Одной из причин развития множества заболеваний, в первую очередь злокачественных, является повышенная экспрессия фактора роста эндотелия сосудов (Vascular Endothelial Growth Factor, VEGF). Бевацизумаб является препаратом для нейтрализации биологической активности VEGF и представляет собой рекомбинантные гуманизированные антитела. Его применение снижает васкуляризацию в очагах повышенной экспресии VEGF, что замедляет опухолевый рост, а также способствует восстановлению зрения при ряде офтальмологических заболеваний. Для определения концентрации бевацизумаба в биологических жидкостях человека представлена тест-система на основе иммуноферментного анализа (ИФА).

Цель

Цель. Целью работы является валидация данной тест-системы.

Материалы и методы

Материалы и методы. Бланковые сыворотки добровольцев, раствор бевацизумаба, иммуноферментный анализ, твердофазный «сэндвич»-ИФА-набор, микропланшетный фотометр.

Результаты и обсуждение

Результаты и обсуждение. Определены следующие характеристики тест-системы: нижний предел количественного определения – 2,0 мкг/мл, верхний калибровочный диапазон – до 200 мкг/мл, правильность и прецизионность внутри одной серии и между сериями не превышает 20 %, а общая ошибка метода – 30 %, краткосрочная стабильность для образцов при комнатной температуре – 6 часов, долгосрочная стабильность – 14 дней при –20 °С, возможность трехкратного замораживания/оттаивания образцов, возможность определения образцов с концентрацией выше верхнего калибратора после разбавления в 2 раза.

Заключение

Заключение. Полученные результаты полностью соответствуют международным критериям приемлемости и позволяют использовать тест-систему ИФА производства ООО «НПЦ Пробиотек» для применения в области клинической лабораторной диагностики.

Introduction

Introduction. One of the reasons for the development of many diseases primarily malignant is the increased expression of Vascular Endothelial Growth Factor (VEGF). Bevacizumab is a drug that neutralizes the biological activity of VEGF. The molecular structure of bevacizumab is a recombinant humanized antibody. Its use reduces vascularization in the foci of increased VEGF expression which slows down tumor growth and also helps restore vision in a number of ophthalmic diseases. To determine the concentration of bevacizumab in human biological fluids a test system based on Enzyme-linked Immunosorbent Assay (ELISA) is presented.

Aim

Aim. Aim of this study is the validation of this test system.

Materials and methods

Materials and methods. Blank sera of volunteers, bevacizumab solution, enzyme-linked immunosorbent assay, solid-phase sandwich ELISA kit, microplate photometer.

Results and discussion

Results and discussion. The following characteristics of the test system were determined: the lower limit of quantification is 2.0 mcg/ml, the upper calibration range is up to 200 mcg/ml, the accuracy and precision within one series and between series does not exceed 20 %, and the total error of the method – 30 %, short-term stability for samples at room temperature – 6 hours, long-term stability – 14 days at –20 °C, the ability to freeze/thaw of samples is up to three times, the ability to determine samples with a concentration above the upper calibrator after diluting in 2 times.

Conclusion

Conclusion. The results obtained fully comply with international acceptance criteria and allow the use of the ELISA test system manufactured by LLC "Probiotek" for use in the field of clinical laboratory diagnostics.  

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