不同细胞培养基对CHO细胞株单克隆抗体产生和糖基化的影响。

IF 2 4区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Cytotechnology Pub Date : 2025-06-01 Epub Date: 2025-03-22 DOI:10.1007/s10616-025-00733-7
Jaeweon Lee, Uriel Ortega-Rodriguez, Chikkathur N Madhavarao, Tongzhong Ju, Thomas O'Connor, Muhammad Ashraf, Seongkyu Yoon
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引用次数: 0

摘要

重组单克隆抗体(mAbs)通常是用中国仓鼠卵巢(CHO)细胞制备的,生物反应器中使用的细胞培养基影响蛋白药物的产量和质量属性。2019冠状病毒病大流行暴露了维持蛋白药物不间断生产和质量一致所需试剂(如培养基和原料)供应链的脆弱性。细胞培养基ActiPro™的供应中断,优化产生VRC01, IgG1-κ单抗,从CHO-K1细胞系,有必要寻找替代培养基。VRC01 mAb是高度糖基化的,可以广泛中和多种人类免疫缺陷病毒(HIV)毒株。我们调查了是否可以在生产中使用替代培养基而不影响糖基化等质量属性。在我们的策略中,我们使用了3种不同的市售培养基,进行了两组实验——有和没有培养基补充,Cell boost 7a和Cell boost 7b。通过比较细胞生长、单抗蛋白的体积产生和糖基化模式来确定替代培养基。在基于细胞生长、单克隆抗体生产潜力和糖基化分析的测试培养基中,ActiCHO™P被发现是比EX-CELL®325 PF CHO培养基更好的替代ActiPro™培养基来生产VRC01单克隆抗体。总的来说,这里使用的方法来确定培养基变化对蛋白质治疗属性的影响,可以在产品开发过程中使用,以建立药物制造中的供应链弹性。补充信息:在线版本包含补充资料,可在10.1007/s10616-025-00733-7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。
Effect of different cell culture media on the production and glycosylation of a monoclonal antibody from a CHO cell line.

Recombinant monoclonal antibodies (mAbs) are commonly produced using Chinese hamster ovary (CHO) cells and the cell culture medium used in bioreactors influences the yield and quality attributes of the protein drug products. The COVID 19 pandemic revealed a vulnerability in the supply chain for necessary reagents (such as culture medium and raw material) for maintaining un-interrupted production of protein drugs with consistent quality. The supply interruption for the cell culture medium ActiPro™ optimized for producing VRC01, an IgG1-κ mAb, from a CHO-K1 cell line, necessitated the search for alternate media. VRC01 mAb is highly glycosylated and can broadly neutralize several strains of Human Immunodeficiency Virus (HIV). We investigated to see if an alternate medium can be used in the production without impacting quality attributes like glycosylation. In our strategy, we used 3 different commercially available media, performed two sets of experiments-with and without media supplements, Cell boost 7a and Cell boost 7b. Cell growth, volumetric production of the mAb protein and glycosylation pattern were compared to identify an alternative medium. Among the tested media based on cell growth, mAb production potential and glycosylation analysis, ActiCHO™ P was found to be a better alternate medium to ActiPro™ medium than EX-CELL® 325 PF CHO medium to produce VRC01 mAb. Overall, the approach used here to establish the impact of variation in medium on protein therapeutic attributes may be used during product development to build in supply chain resilience in drug manufacturing.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00733-7.

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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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