Development of a Serum-Free Media Based on the Optimal Combination of Recombinant Protein Additives and Hydrolysates of
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lopment of a Serum-Free Media Based on the Optimal Combination of Recombinant Protein Additives and Hydrolysates of Non-Animal Origin to Produce Immunoglobulins D. S. Balabashina, *, **, ***, E. N. Kaliberdaa, I. V. Smirnova, Y. A. Mokrushinaa, T. V. Bobika, T. K. Alieva, b, D. A. Dolgikha, c, and M. P. Kirpichnikova, c a
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, 117997 Russia b Chemistry Department, Moscow State University, Moscow, 119991 Russia c Biology Department, Moscow State University, Moscow, 119991 Russia *e-mail: [email protected] **e-mail: [email protected] ***e-mail: [email protected] Received December 30, 2019; revised April 14, 2020; accepted April 22, 2020
Abstract—The potential ability to cultivate a recombinant immunoglobulin (IgG1)-producer line in Iscove’s Modified Dulbecco Medium (IMDM) as the base cultivation medium with the introduction of recombinant proteins and hydrolysates of non-animal origin was studied. Recombinant insulin, human serum albumin (produced by E. coli bacteria), and transferrin (by Pichia pastoris yeasts) were used to enrich the medium. A combination of protein additives (part of the developed medium) allowed an increase in the productivity of a stable producer culture of recombinant humanized antibodies based on CHO-line cells by 44% as compared with the used complex commercial insulin–transferrin–selenium (ITS) additive. The introduction of pea and rice protein hydrolysates containing peptides with a molecular weight lower than 5 kDa to modified IMDM medium contributed to an increase in line productivity by 3.9 and 4.5 times, respectively. Keywords: plant protein hydrolysates, yeast extract, insulin, transferrin, sodium selenite, HSA, antibodies, CHO DG44, stable cell line DOI: 10.1134/S0003683820050038
INTRODUCTION Recombinant therapeutic proteins expanded the possibilities of modern medicine, allowing the development of new and efficient methods of treatment for many diseases, from cancer to infertility [1]. Approximately 60–70% of all recombinant pharmaceutical proteins are obtained from mammalian cells. Many of these proteins are expressed in immortalized Chinese hamster ovary (CHO) cells, which make it possible to preserve, as much as possible, not only their structure but also their function. Thus, they can be used in many fields of medicine, biotechnology, and agriculture. Other eukaryotic cell lines, such as mouse myeloma (NSO), baby hamster kidney (BHK) cells, and human embryonic kidney cells (HEK-293), are also used in the production of recombinant proteins. However, they have a narrow specificity and direction of action; in this regard, their use is strictly limited by the area of application [2]. CHO-line cells are becoming an increasingly important tool for the production of recombinant therapeutic proteins due to their rapid growth, the stability of the expression of foreign genes
in them, and the high yield of recombinant proteins [3]. An increase in the yield of a recombinant protein and, consequently
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