Novel application of anti-human Fc nanobody for screening high-producing CHO cells for monoclonal antibody

IF 3.9 4区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Engineering in Life Sciences Pub Date : 2022-08-23 DOI:10.1002/elsc.202200028
Di Zhu, Zheng Wang, Yunxia Xu, Jing Lin, Mei Qiu, Jianghai Liu, Xinlei Li
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Abstract

Animal-derived anti-IgG secondary antibodies are currently employed to stain and screen of human monoclonal antibody(mAb)-producing cells, but using animal-derived antibodies may raise the concerns of high cost, complicated operations and biological safety issues in biopharmaceutical manufacturing. Nanobodies(VHHs) are attractive forms of antibodies for their straightforward engineering and expression in both eukaryotic and prokaryotic systems. Using phage-displayed immune llama VHH library, we identified new anti-Fc VHHs that could bind to human Fc with high affinity. In GFP fusion format, the anti-Fc VHH-GFP generated dramatically stronger FACS signals than AF488 conjugated anti-IgG antibodies when used for staining mAb-producing CHO cells. Furthermore, preparative sorting of CHO cells based on anti-Fc VHH-GFP staining resulted in the enrichment of cell lines capable of synthesizing mAb at high productivity. This safe and cost-efficient anti-Fc VHH-GFP may optimize the process of generating highly productive cell lines for therapeutic mAb production compared to conventional animal-derived fluorescent antibodies.

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抗人Fc纳米体在筛选高产CHO细胞单克隆抗体中的新应用
动物源性抗igg二抗目前用于人单克隆抗体(mAb)产生细胞的染色和筛选,但在生物制药生产中,使用动物源性抗体可能会引起成本高、操作复杂和生物安全性问题的担忧。纳米小体(VHHs)是一种有吸引力的抗体形式,因为它们在真核和原核系统中都具有简单的工程和表达。利用噬菌体展示的免疫羊驼vhh文库,我们发现了新的抗Fc VHH,可以高亲和力地与人类Fc结合。在GFP融合格式中,当用于染色产生单克隆抗体的CHO细胞时,抗fc VHH-GFP比AF488结合的抗igg抗体产生更强的FACS信号。此外,基于抗fc VHH-GFP染色对CHO细胞进行制备分选,富集了能够高产量合成单抗的细胞系。与传统的动物源性荧光抗体相比,这种安全且经济高效的抗fc VHH-GFP可以优化生产用于治疗性单抗生产的高产细胞系的过程。
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来源期刊
Engineering in Life Sciences
Engineering in Life Sciences 工程技术-生物工程与应用微生物
CiteScore
6.40
自引率
3.70%
发文量
81
审稿时长
3 months
期刊介绍: Engineering in Life Sciences (ELS) focuses on engineering principles and innovations in life sciences and biotechnology. Life sciences and biotechnology covered in ELS encompass the use of biomolecules (e.g. proteins/enzymes), cells (microbial, plant and mammalian origins) and biomaterials for biosynthesis, biotransformation, cell-based treatment and bio-based solutions in industrial and pharmaceutical biotechnologies as well as in biomedicine. ELS especially aims to promote interdisciplinary collaborations among biologists, biotechnologists and engineers for quantitative understanding and holistic engineering (design-built-test) of biological parts and processes in the different application areas.
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