各种双糖适应对CHO-K1悬浮细胞重组IgA1产生的影响。

IF 2 4区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
John Benson D Choa, Tadahiro Sasaki, Hiroyuki Kajiura, Kazuyoshi Ikuta, Kazuhito Fujiyama, Ryo Misaki
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引用次数: 0

摘要

免疫球蛋白A (IgA)作为一种新的治疗性抗体已显示出潜力。然而,重组IgA存在产量低的问题。补充培养基是提高重组蛋白产量和质量的有效途径。在本研究中,我们将产生IgA1的CHO-K1悬浮细胞适应于高浓度(150 mM)的不同双糖,即蔗糖、麦芽糖、乳糖和海藻糖,以提高重组IgA1的产量和质量。适应二糖的细胞系细胞生长速度较慢,但与不适应iga1的细胞系相比,它们的细胞活力得到了延长。除了适应麦芽糖的细胞系外,所有细胞系的葡萄糖消耗都耗尽了,即使在培养的第9天,它仍然含有葡萄糖。乳酸产量在适应双糖的细胞系中较高。适应麦芽糖的iga1生产系的比生产力是不适应的4.5倍。此外,这种特异性生产力比以前的含有lambda链的重组IgA1产品更高。最后,分泌的IgA1在所有细胞系中聚集,这可能是由自聚集引起的。这种聚集也在麦芽糖适应细胞系的细胞内开始。这些结果表明,高浓度的双糖补充诱导了产生iga1的CHO-K1细胞系的高渗透压。此外,适应麦芽糖的CHO-K1细胞系受益于额外的碳水化合物来源。补充信息:在线版本包含补充资料,可在10.1007/s10616-023-00571-5获得。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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

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

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

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

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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来源期刊
Cytotechnology
Cytotechnology 生物-生物工程与应用微生物
CiteScore
4.10
自引率
0.00%
发文量
49
审稿时长
6-12 weeks
期刊介绍: The scope of the Journal includes: 1. The derivation, genetic modification and characterization of cell lines, genetic and phenotypic regulation, control of cellular metabolism, cell physiology and biochemistry related to cell function, performance and expression of cell products. 2. Cell culture techniques, substrates, environmental requirements and optimization, cloning, hybridization and molecular biology, including genomic and proteomic tools. 3. Cell culture systems, processes, reactors, scale-up, and industrial production. Descriptions of the design or construction of equipment, media or quality control procedures, that are ancillary to cellular research. 4. The application of animal/human cells in research in the field of stem cell research including maintenance of stemness, differentiation, genetics, and senescence, cancer research, research in immunology, as well as applications in tissue engineering and gene therapy. 5. The use of cell cultures as a substrate for bioassays, biomedical applications and in particular as a replacement for animal models.
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