基于紫外-可见光谱的CHO细胞培养液快速分析方法  

Rapid analysis of CHO cell culture medium based on UV-Vis spectroscopy

作　　者：阎续 万宇翔 聂磊 王海彬 瞿海斌[1] YAN Xu;WAN Yu-xiang;NIE Lei;WANG Hai-bin;QU Hai-bin(Pharmaceutical Informatics Institute,Zhejiang University,Hangzhou 310058,China;Hisun Biopharmaceutical Co.Ltd.,Hangzhou 311404,China)

机构地区：[1]浙江大学药物信息学研究所,浙江杭州310058 [2]海正生物制药有限公司,浙江杭州311404

出　　处：《高校化学工程学报》2021年第5期860-866,共7页Journal of Chemical Engineering of Chinese Universities

基　　金：浙江省科技计划项目重点研发计划(2017C03003)。

摘　　要：为了建立基于紫外-可见(UV-Vis)光谱和酚红的快速分析方法,用于同时测定细胞培养液中葡萄糖(Gluc)和乳酸(Lac)质量浓度、谷氨酸(Glu)和谷氨酰胺(Gln)浓度及活细胞密度(VCD)、总细胞密度(TCD)、细胞活度(Viability)和单克隆抗体表达量(Titer),将细胞培养液上清与酚红溶液混合,采集混合溶液的UV-Vis光谱,以偏最小二乘(PLS)法建立UV-Vis光谱与各指标的多元校正模型。发现Lac质量浓度、Glu浓度、VCD、TCD、Viability和Titer模型预测性能较好;Gluc质量浓度和Gln浓度模型预测性能较差。结果表明,将该方法应用于细胞培养过程多指标快速分析具有一定可行性,为生产过程实时监控提供了研究基础。A rapid analytical method based on UV-Vis spectroscopy and phenol red was developed for simultaneous quantitative determination of mass concentrations of glucose(Gluc) and lactate(Lac),concentrations of glutamate(Glu) and glutamine(Gln), viable cell density(VCD), total cell density(TCD),cell viability(Viability) and monoclonal antibody(mAb) titer(Titer) in cell culture medium. UV-Vis spectra of mixtures of cell culture supernatants and phenol red solution were collected. Partial least squares(PLS)was used to develop multivariate calibration models between UV-Vis spectra and parameters. The results show that models of Lac mass concentration, Glu concentration, VCD, TCD, Viability and Titer have satisfactory predictive capacity, while those of Gluc mass concentration and Gln concentration have unsatisfactory predictive capacity. This study presents the feasibility of applying the proposed method for rapid analysis of multiple parameters in cell culture, and provides the basis for real time monitoring and control for manufacturing.

关 键 词：紫外-可见光谱 酚红 偏最小二乘 CHO细胞培养 快速分析 单克隆抗体 

分 类 号：R917[医药卫生—药物分析学]