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      Effects of various disaccharide adaptations on recombinant IgA1 production in CHO-K1 suspension cells

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          Abstract

          Immunoglobulin A (IgA) has been showing potential as a new therapeutic antibody. However, recombinant IgA suffers from low yield. Supplementation of the medium is an effective approach to improving the production and quality of recombinant proteins. In this study, we adapted IgA1-producing CHO-K1 suspension cells to a high concentration (150 mM) of different disaccharides, namely sucrose, maltose, lactose, and trehalose, to improve the production and quality of recombinant IgA1. The disaccharide-adapted cell lines had slower cell growth rates, but their cell viability was extended compared to the nonadapted IgA1-producing cell line. Glucose consumption was exhausted in all cell lines except for the maltose-adapted one, which still contained glucose even after the 9th day of culturing. Lactate production was higher among the disaccharide-adapted cell lines. The specific productivity of the maltose-adapted IgA1-producing line was 4.5-fold that of the nonadapted line. In addition, this specific productivity was higher than in previous productions of recombinant IgA1 with a lambda chain. Lastly, secreted IgA1 aggregated in all cell lines, which may have been caused by self-aggregation. This aggregation was also found to begin inside the cells for maltose-adapted cell line. These results suggest that a high concentration of disaccharide-supplemented induced hyperosmolarity in the IgA1-producing CHO-K1 cell lines. In addition, the maltose-adapted CHO-K1 cell line benefited from having an additional source of carbohydrate.

          Supplementary Information

          The online version contains supplementary material available at 10.1007/s10616-023-00571-5.

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          Biopharmaceutical benchmarks 2018

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            CHO cells in biotechnology for production of recombinant proteins: current state and further potential.

            Recombinant Chinese hamster ovary cells (rCHO) cells have been the most commonly used mammalian host for large-scale commercial production of therapeutic proteins. Recent advances in cell culture technology for rCHO cells have achieved significant improvement in protein production leading to titer of more than 10 g/L to meet the huge demand from market needs. This achievement is associated with progression in the establishment of high and stable producer and the optimization of culture process including media development. In this review article, we focus on current strategies and achievements in cell line development, mainly in vector engineering and cell engineering, for high and stable protein production in rCHO cells. The approaches that manipulate various DNA elements for gene targeting by site-specific integration and cis-acting elements to augment and stabilize gene expression are reviewed here. The genetic modulation strategy by "direct" cell engineering with growth-promoting and/or productivity-enhancing factors and omics-based approaches involved in transcriptomics, proteomics, and metabolomics to pursue cell engineering are also presented.
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              Is Open Access

              A cross-reactive human IgA monoclonal antibody blocks SARS-CoV-2 spike-ACE2 interaction

              COVID-19 caused by SARS-CoV-2 has become a global pandemic requiring the development of interventions for the prevention or treatment to curtail mortality and morbidity. No vaccine to boost mucosal immunity, or as a therapeutic, has yet been developed to SARS-CoV-2. In this study, we discover and characterize a cross-reactive human IgA monoclonal antibody, MAb362. MAb362 binds to both SARS-CoV and SARS-CoV-2 spike proteins and competitively blocks ACE2 receptor binding, by overlapping the ACE2 structural binding epitope. Furthermore, MAb362 IgA neutralizes both pseudotyped SARS-CoV and SARS-CoV-2 in 293 cells expressing ACE2. When converted to secretory IgA, MAb326 also neutralizes authentic SARS-CoV-2 virus while the IgG isotype shows no neutralization. Our results suggest that SARS-CoV-2 specific IgA antibodies, such as MAb362, may provide effective immunity against SARS-CoV-2 by inducing mucosal immunity within the respiratory system, a potentially critical feature of an effective vaccine.
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                Author and article information

                Contributors
                misaki@icb.osaka-u.ac.jp
                Journal
                Cytotechnology
                Cytotechnology
                Cytotechnology
                Springer Netherlands (Dordrecht )
                0920-9069
                1573-0778
                16 March 2023
                : 1-11
                Affiliations
                [1 ]GRID grid.136593.b, ISNI 0000 0004 0373 3971, International Center for Biotechnology, , Osaka University, ; 2-1 Yamada-Oka, Suita, Osaka 565-0871 Japan
                [2 ]GRID grid.136593.b, ISNI 0000 0004 0373 3971, Department of Viral Infections, Research Institute for Microbial Diseases, , Osaka University, ; 3-1 Yamada-Oka, Suita, Osaka 565-0871 Japan
                [3 ]GRID grid.136593.b, ISNI 0000 0004 0373 3971, Institute for Open and Transdisciplinary Research Initiatives (OTRI), , Osaka University, ; 2-1 Yamada-Oka, Suita, Osaka 565-0871 Japan
                [4 ]GRID grid.136593.b, ISNI 0000 0004 0373 3971, Research Institute for Microbial Diseases, , Osaka University, ; 3-1 Yamada-Oka, Suita, Osaka 565-0871 Japan
                [5 ]BioAcademia, Inc, 3-1 Yamada-Oka, Suita, Osaka 565-0871 Japan
                [6 ]GRID grid.10223.32, ISNI 0000 0004 1937 0490, Faculty of Science, Osaka University Cooperative Research Station in Southeast Asia (OU:CRS), , Mahidol University, ; Bangkok, Thailand
                Article
                571
                10.1007/s10616-023-00571-5
                10018586
                8eac17f4-2186-4d59-a574-e52034bc9aed
                © The Author(s), under exclusive licence to Springer Nature B.V. 2023, Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

                This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.

                History
                : 5 October 2022
                : 9 February 2023
                Funding
                Funded by: The Japan Science and Technology Agency/Japan International Cooperation Agency, Science and Technology Research Partnership for Sustainable Development (JST/JICA, SATREPS)
                Award ID: 08080924
                Award ID: 08080924
                Categories
                Research

                Biotechnology
                chinese hamster ovary cell,disaccharide,immunoglobulin a,productivity
                Biotechnology
                chinese hamster ovary cell, disaccharide, immunoglobulin a, productivity

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