高通量筛选在双特异性抗体亲和层析工艺开发中的应用  

High Throughput Screening in Affinity Chromatography Process Development of Bispecific Antibody

作　　者：梁晓莹 李谦 何清泉 王子臣 赵鸿阳 李国柱 秦国宏 LIANG Xiaoying;LI Qian;HE Qingquan;WANG Zichen;ZHAO Hongyang;LI Guozhu;QIN Guohong(Fanghua Pharmaceutical Research Institute,Nanjing Chia-Tai Tianqing Pharmaceutical Co.,Ltd.,Nanjing 210046,China)

机构地区：[1]南京正大天晴制药有限公司芳华药物研究院,江苏南京210046

出　　处：《生物化工》2025年第1期102-107,127,共7页Biological Chemical Engineering

摘　　要：目的:为快速评估填料及筛选工艺条件,探究高通量筛选技术在双特异性抗体的亲和层析工艺开发中的应用。方法:采用高通量微孔滤板进行工艺开发,利用Langmuir等温吸附曲线评估8种商业Protein A填料的结合能力,筛选出高载量的2种填料后对5种洗脱条件和7种淋洗条件进行筛选,并在层析柱上对确定的工艺进行验证。结果:筛选出的BioCap rProtein A PC填料以60 g/L的载量,在pH=3.6的洗脱溶液和含0.5 mol/L NaCl的淋洗2的工艺下,柱层析的收率(亲和收率和中和收率分别为93.6%和100.0%)及产品质量(纯度95.5%,宿主细胞蛋白含量1282 ng/mg)均满足工艺要求。结论:高通量筛选技术可快速、高效地从多种亲和填料中筛选出最佳填料以及从多种工艺参数中筛选出适合的工艺条件,缩短了实验周期,提高了实验效率。Objective:The application of high-throughput screening technology in developing affinity chromatography for bispecific antibodies is explored to evaluate the packing material and rapidly screen the process conditions.Methods:The process is developed using high-throughput microwell filter plates,the binding capacity of 8 commercial Protein A resins is evaluated by using Langmuir isothermal adsorption curves,after screening out 2 kinds of fillers with high loading,5 elution conditions and 7 elution conditions are screened,and the determined process is verified on chromatography columns.Results:The screened BioCap rProtein A PC resin,with a loading capacity of 60 g/L,is employed in a process utilizing an elution solution of pH=3.6 and a wash 2 containing with 0.5 mol/L NaCl.The yield of column chromatography(affinity yield and neutralization yield are 93.6%and 100.0%,respectively)and product quality(purity 95.5%,host cell protein content 1282 ng/mg)meet the process requirements.Conclusion:The high-throughput screening technology can quickly and efficiently screen the best resin from a variety of affinity resins and select the appropriate process conditions from a variety of process parameters,which shortens the experimental cycle and improves the experimental efficiency.

关 键 词：高通量筛选 亲和层析 双特异性抗体 

分 类 号：TQ464.7[化学工程—制药化工]