Effects of Fibroblast Growth Factor-2 and Other Microsupplements on the Productivity of IgG- and IgA-Producing Cell Line
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OGICALS TECHNOLOGY
Effects of Fibroblast Growth Factor-2 and Other Microsupplements on the Productivity of IgG- and IgA-Producing Cell Lines V. V. Argentovaa, *, T. K. Alievb, M. E. Gasparyanc, D. A. Dolgikha, c, and M. P. Kirpichnikova, c aBiology
Department, Lomonosov Moscow State University, Moscow, 119991 Russia Department, Lomonosov Moscow State University, Moscow, 119991 Russia c Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, 117997 Russia *e-mail: [email protected] bChemistry
Received August 12, 2019; revised October 24, 2019; accepted February 11, 2020
Abstract—Several serum-free media have been developed and are successfully used at present for the cultivation of various recombinant eukaryotic cell lines. Therapeutic recombinant IgA antibodies have recently been increasingly used along with numerous IgG1 isotype antibodies. Therefore, it became necessary to improve the growth characteristics, metabolism, and productivity of cell cultures producing these antibodies. The stimulating effect of a complex supplement containing Zn salts and FGF-2 on cell cultivation on basal media, DMEM and IMDM, was revealed in this work. The addition of dextran sulfate sodium salt and iron citrate to the basal medium was accompanied by improved productivity of stable IgG- and IgA-producing cell lines, as well as the homogeneity and density of cell cultures, which is most important in IgA-antibody production. Keywords: serum-free media, microsupplements, recombinant antibodies, fibroblast growth factor-2 DOI: 10.1134/S0003683820090021
INTRODUCTION The growing industrial demand for therapeutic antibodies is driving research to increase the productivity of mammalian cell lines. The development and improvement of serum-free medium compositions and supplements to them have become one of the main trends in the optimization of therapeutic antibody production. They are aimed not only at an increase in cell line productivity but also at improvement of the quality of the produced recombinant immunoglobulins and prevention of the risk of contamination by animal components, which are potential sources of various pathogens. However, most mammalian cells are extremely sensitive both to changes in culturing conditions and to the number and quality of media supplements. The effective cultivation of mammalian cells requires the presence of all nutrient compounds, including amino acids, vitamins, and inorganic additives [1, 2]. It is known that vitamins and amino acids added to protein-free or serum-free media significantly increase the density of cell cultures and their antibody production [1, 3, 4]. An increase in the production of Abbreviations: ACR—anticlumping reagent; CHO—Chinese hamster ovarian cells; DMEM/F12—medium consisting of DMEM and F12 media; ELISA—enzyme-linked immunosorbent assay; FGF-2—fibroblast growth factor 2; IMDM— Iscove’s modified Dulbecco’s medium; ITS—insulin-transferrin-selenite supplement; PBS—phosphate-buffered saline.
therapeutic proteins and improved cell grow
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