Abstract
The Her2 receptor is an important target for antitumor therapy in the treatment of breast cancer. Trastuzumab, based on anti-Her2 monoclonal antibodies, is used in clinical practice. Trastuzumab is produced by animal cells culture technology and is quite expensive. We use the technology of production of recombinant antibodies in the plants Nicotiana benthamiana with a high yield of final purified protein. Objective. The aim of following study is a comparison of monoclonal antibodies received via classic cell culture technology and produced in plant biomass. Materials and methods. Recombinant plant-made antibodies were isolated by affinity chromatography from the biomass of N. benthamiana plants agroinfiltrated by vector constructs. Comparison of affinity properties was carried out by immunocytochemical staining of cells and competitive binding using flow cytometry analysis. Results and conclusion. We show that the antibodies expressed in N. benthamiana are equal to those obtained from mammalian cells in binding to Her2 antigen localized on the surface of the SK-BR-3 cells. In the present work it was shown that the plant-made anti-Her2 antibodies do not differ in specific binding with the Her2 antigen, as well as with IV subdomain of the Her2 receptor.
Highlights
The Her2 receptor is an important target for antitumor therapy in the treatment of breast cancer
Recombinant plant-made antibodies were isolated by affinity chromatography from the biomass of N. bentha miana plants agroinfiltrated by vector constructs
We show that the antibodies expressed in N. benthamiana are equal to those obtained from mammalian cells in binding to Her2 antigen localized on the surface of the SK-BR-3 cells
Summary
Блохина» Минздрава России; Россия, 115478 Москва, Каширское шоссе, 24; 2ФГБУН Институт общей генетики им. В настоящее время в клинической практике применяют препарат на основе моноклональных антител анти-Her2 – трастузумаб. Трастузумаб производится в культуре животных клеток и стоит достаточно дорого. Разработана технология продукции рекомбинантных антител в растительном продуценте Nicotiana benthamiana с высоким выходом очищенного белка. Цель исследования – сравнение аффинных свойств рекомбинантных антител, полученных классическим способом в культуре клеток и полученных в растительной биомассе. Сравнение аффинных свойств проводили методами иммуноцитохимической окраски клеток, конкурентного связывания нативным эпитопом с анализом методом проточной цитофлуориметрии. В данной работе нами установлено, что антитела, экспрессированные в N. benthamiana, не уступают аналогам, полученным из клеток млекопитающих, в связывании антигена Her на поверхности мембран клеток SK-BR-3. Ключевые слова: рак молочной железы, терапевтические антитела, продукция антител в растениях.
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