抗ErbB2嵌合抗体chA21大规模纯化工艺的建立及质量研究  

Mass-scale Purification and Quality Control of a Single-chain Chimeric Anti-ErbB2 Antibody

作　　者：傅璐[1] 黄辉 陈舒扬[1] 初波[1] 刘兢[1] 

机构地区：[1]中国科学技术大学生命科学学院,合肥230027 [2]安科生物(集团)股份有限公司,合肥230081

出　　处：《中国生物工程杂志》2009年第7期68-73,共6页China Biotechnology

基　　金：国家自然科学基金(30570362);教育部博士点基金(20060358021);国家"863"计划重点项目子课题(2006AA02A245)资助项目

摘　　要：自行研制的抗ErbB2嵌合抗体chA21具有抑制高表达ErbB2的乳腺癌细胞生长的作用。在前期小规模培养和纯化工作的基础上,以填充床生物反应器大规模培养CHO工程细胞株表达的上清为原料,采用亲和层析、凝胶过滤除盐、阳离子交换层析、分子筛等步骤,分离纯化嵌合抗体chA21,建立了大规模纯化工艺,并按照中国药典(2005年版第三部)对最终产品进行全面鉴定和质量控制。该工艺能有效解决抗体纯化过程形成的多聚体问题,去除内毒素和DNA残留;可以确保每批纯化20～40L培养上清,每批收获嵌合抗体可达5g以上,蛋白总回收率大于50%,纯度可达98%。研究结果表明,该抗体纯化工艺得率高,质量控制方法稳定可靠,适用于大规模生产。Single-chain Chimeric Anti-ErbB2 Antibody chA21 was constructed in our libratory, which could be a drug candidate for breast cancer therapy, chA21 from engineered CHO cells were cultured in the packed bed bioreactor. To establish mass-scale purification technology, culture supernatant was collected and purified in a sequence of downstream processing steps, namely, affinity chromatography, desahing, ion exchange chromatography and molecular sieve chromatography; and its quality was all analyzed according to the Chinese Pharmacopoeia （Volume Ⅲ, 2005 Edition）. By the developed mass-scale procedure, formed aggregates could be removed effetely; endotoxins and residual DNA were also extracted. 20 -40L of culture superuatant could be treated in a single operation and more than 5g of chA21 was purified. The purify rate reached more than 98% and the total recovery rate was over 50%. So the purify rate of the produce technics was high and quality inspection methods were stability and reliable, which could be suitable in large-scale production of chA21.

关 键 词：嵌合抗体 纯化工艺 质量控制 聚集 

分 类 号：Q819[生物学—生物工程]