摘要
目的对WuT3杂交瘤细胞生物反应器无血清培养上清中单克隆抗体进行分离纯化,建立中试分离纯化工艺。方法采用两步离子交换层析法结合硫酸铵盐析法分离纯化WuT3单抗。在通过小量试验优化纯化参数后,进行中量试验和中试放大。结果建立的纯化方法可以放大到中试规模,每批投料30000ml培养上清,可收获单抗达1.5g,单抗总回收率大于60%。经分离纯化后,单抗IgG纯度大于95%,采用免疫荧光法检测抗体活性合格。结论建立了一条生物反应器无血清培养杂交瘤细胞大规模分离纯化单抗的工艺路线。
Objective To develop a pilot procedure for purification of monoclonal antibody （McAb） from serum-free culture superuatant of WuT3 hybridoma cells in bioreactor. Methods Purify WuT3 McAb by two-step ion exchange chromatography combined with ammonium sulfate precipitation. After the condition for purification was optimized, the purification procedure was scaled up. On the basis of this, a pilot purification procedure was developed. Results By the developed pilot procedure,30 000 ml of culture supematant was treated in a single operation,from which 1.5 g of McAb was purified. The total recovery rate of McAb was more than 60%. The pu- rified MeAb reached a purity of more than 95% and showed immunological activity as proved by IFA. Conclusion A pilot procedure for purification of McAb from serum-free culture of hybridoma cells in bioreactor was developed.
出处
《中国生物制品学杂志》
CAS
CSCD
2007年第8期593-595,599,共4页
Chinese Journal of Biologicals
基金
国家重点科技攻关项目96-C02-03-05部分工作.
关键词
无血清培养
杂交瘤细胞
单克隆抗体
纯化工艺
Serum-free culture
Hybridoma cells
Monoclonal antibody
Purification procedure
