Abstract
Quality control of recombinant interferon (rIFN) products with the help of modern analytical methods, including those used for identification, is becoming increasingly relevant nowadays. Identification is especially challenging in the case of Russian rIFN products that contain not only interferon (IFN) alpha-2b, but also other active ingredients and excipients that hinder the use of physical and chemical methods. Manufacturers of such products use IFN neutralization assay with mono- and/or polyclonal antibodies for identification. The aim of the study was to assess the feasibility of using different types of antibodies in the identification test based on neutralization of IFN antiviral activity in IFN alpha-2b products containing other active ingredients and excipients in addition to IFN. Materials and methods: the following materials were used in the study: MDBK cells, vesicular stomatitis virus, samples of IFN alpha-2b products with different composition and by different manufacturers, mono- and polyclonal antibodies by different manufacturers. Identification of rINFs was carried out by a biological method based on neutralization by specific antibodies of IFN ability to suppress the cytopathic effect of the indicator virus in a cell culture using a reference standard for comparison. Results: both polyclonal and monoclonal antibodies were shown to neutralize the activity of the tested IFN alpha-2b substances. Polyclonal antibodies interact with all products containing the same active ingredients, irrespective of their composition. Monoclonal antibodies interact selectively with some products. Conclusions: polyclonal antibodies can be used for identification of any product containing IFN alpha-2b. The use of monoclonal antibodies for this purpose is limited and depends on the composition of the product.
Highlights
Сравнительная оценка использования моно- и поликлональных антител при определении подлинности различных лекарственных средств на основе интерферона альфа-2b
Comparative evaluation of mono- and polyclonal antibodies used in identification of interferon alpha-2b products
Quality control of recombinant interferon products with the help of modern analytical methods, including those used for identification, is becoming increasingly relevant nowadays
Summary
No MAB411, клон ST2541 (далее — АТ МАВ411) и Mouse Anti-Human α-Interferon Monoclonal Antibody, кат. Приготовленные смеси и контрольные образцы инкубировали при температуре (37±1) °C в течение 1 ч, после чего из лунок 96‐луночного планшета с клеточным монослоем удаляли культуральную среду и вносили (в 4 раздельные лунки на каждое разведение антител) нейтрализованные смеси РРЛС и РР-МСО (объемом по 100 мкл). Для контроля активности интерферона вносили в лунки по 100 мкл КО-МСО и КО-ЛС (по 4 лунки на каждый контрольный образец). После внесения растворов 96‐луночный планшет инкубировали в течение 22–26 ч в стандартных условиях, а затем из лунок удаляли культуральную среду и во все лунки, содержащие нейтрализованные смеси и контрольные образцы, вносили по 100 мкл раствора индикаторного вируса в рабочей дозе (титр и рабочую дозу вируса определяли в ходе определения специфической активности). Лекарственное средство выдерживало испытание на подлинность, если нейтрализация активности РР-ЛС и РР-МСО происходила при аналогичных разведениях антител
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