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A single-step high-throughput bioassay for quantifying Fc-containing recombinant proteins based on non-classical calculation of fluorescence polarization

文献类型：期刊论文

作者Zheng, Yujuan2,4; Chen, Ganjun3; Liu, Guojian1; Rana, Gul E.2,4; Wang, Chunhe2,3,4
刊名ANALYTICAL METHODS
出版日期2024-06-20
卷号16期号:24页码:3917-3926
ISSN号1759-9660
DOI10.1039/d4ay00372a
通讯作者Wang, Chunhe()
英文摘要The titer of recombinant proteins is one of the key parameters in biopharmaceutical manufacturing processes. The fluorescence polarization (FP)-based assay, a homogeneous, high-throughput and real-time analytical method, had emerged as a powerful tool for biochemical analysis and environmental monitoring. In this study, an FP-based bioassay was utilized to quantify antibody fragment crystallizable (Fc)-containing proteins, such as recombinant monoclonal antibodies (mAbs) and mAb derivatives, in the cell culture supernatant, and the impacts of tracer molecular weight and FITC-coupling conditions on fluorescence polarization were methodically examined. Distinct from the fluorescence polarization potency calculated by classical formula, we for the first time proposed a new concept and calculation of fluorescence polarization intensity, based on which an analytical method with broader detection range and analysis window was established for quantifying Fc-containing proteins. This provided new ideas for the practical application of fluorescence polarization theory. The established method could detect 96 samples within 30 minutes, with dynamic titer range of 2.5-400 mg L-1, and a linear fitting R2 between the measured and actual concentration reaching 0.99. The method had great application prospects in determining the titer of recombinant proteins with Fc fragments, especially when applied to large-scale screening of high-yield and stable expression CHO cell lines commonly used in biopharmaceutical industry. A single-step and high-throughput bioassay based on non-classical calculation of fluorescence polarization for quantifying Fc-containing recombinant proteins in the cell culture supernatant.
WOS关键词ANTIBODY ; FUSION ; PROBES
资助项目Special Project for Research and Development in Key Areas of Guangdong Province[2022B1111070007] ; Key-Area Research and Development Program of Guangdong Province[32370958] ; Key-Area Research and Development Program of Guangdong Province[81872785] ; National Natural Science Foundation of China[LX211005] ; CAS Bohai Rim Advanced Research Institute for Drug Discovery Project[210205143867019] ; CAS Bohai Rim Advanced Research Institute for Drug Discovery Project[191022172638719] ; Major Scientific and Technological Special Project of Zhongshan City
WOS研究方向Chemistry ; Food Science & Technology ; Spectroscopy
语种英语
WOS记录号WOS:001237962300001
出版者ROYAL SOC CHEMISTRY
源URL[http://119.78.100.183/handle/2S10ELR8/311576]  
专题中国科学院上海药物研究所
通讯作者Wang, Chunhe
作者单位1.Shanghai Mabstone Biotechnol Co Ltd, Shanghai 201203, Peoples R China
2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
3.Dartsbio Pharmaceut Co Ltd, Zhongshan 528400, Peoples R China
4.Chinese Acad Sci, Shanghai Inst Mat Med, Biotherapeut Discovery Res Ctr, Shanghai 201203, Peoples R China
推荐引用方式
GB/T 7714		 Zheng, Yujuan,Chen, Ganjun,Liu, Guojian,et al. A single-step high-throughput bioassay for quantifying Fc-containing recombinant proteins based on non-classical calculation of fluorescence polarization[J]. ANALYTICAL METHODS,2024,16(24):3917-3926.
APA Zheng, Yujuan,Chen, Ganjun,Liu, Guojian,Rana, Gul E.,&Wang, Chunhe.(2024).A single-step high-throughput bioassay for quantifying Fc-containing recombinant proteins based on non-classical calculation of fluorescence polarization.ANALYTICAL METHODS,16(24),3917-3926.
MLA Zheng, Yujuan,et al."A single-step high-throughput bioassay for quantifying Fc-containing recombinant proteins based on non-classical calculation of fluorescence polarization".ANALYTICAL METHODS 16.24(2024):3917-3926.

入库方式： OAI收割

来源：上海药物研究所

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