|
|
|
| Mass-scale purification and quality control of a single-chain chimeric Anti-ErbB2 antibody |
|
|
|
Abstract Abstract 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, desalting, ion exchange chromatography and molecular sieve chromatography; and its quality was all analyzed according to the Chinese Pharmacopoeia (VolumeⅢ, 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 supernatant 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.
|
|
Received: 08 April 2009
Published: 28 July 2009
|
|
|
| Fund: National Nature Science Foundation of China;specialized Research Fund for the Doctoral Program of Higher Education;Hi-Tech Research and Development Program (‘‘863’’ Program) of China |
|
Corresponding Authors:
Jing liu
|
|
|
| ［1］ Jukkola A, Bloigu R, Soini Y, et al. c-erbB-2 positivity is a factor for poor prognosis in breast cancer and poor response to hormonal or chemotherapy treatment in advanced disease. Eur J Cancer, 2001, 37(3):347~354
［2］ Slamon D J, Godolphin W, Jones L A, et al. Studies of the HER2 /neu proto-oncogene in human breast and ovarian cancer. Science, 1989, 244 (4905): 707~712
［3］ Goldenberg M M. Trastuzumab, a recombinant DNA-derived humanized monoclonal antibody, a novel agent for the treatment of metastatic breast cancer. Clin Ther. 1999, 21(2):309~318
［4］ Cheng L S, Liu A P, Yang J H, et al. Construction, expression and characterization of the engineered antibody against tumor surface antigen, p185 (c-erbB-2). Cell Res, 2003, 13(1): 35~48
［5］ Wang J, Shi Y, Liu Y, et a1. Purification characterization of a single-chain chimeric anti-p185 antibody expressed by CHO-GS system. Protein Expr Purif, 2005, 41(1): 68~76
［6］ 马军, 董艳秋, 邹珉, 等. 丁酸钠和丙酸钠对工程CHO细胞生长代谢和嵌合抗体表达的影响. 中国生物工程杂志, 2005, 25(10):12~16
Ma J, Dong Y Q, Zou M, et a1. China Biotechnology, 2005, 25(10): l2~16
［7］ 乔婧娟, 刘海波, 朱娟娟, 等. 抗P185工程抗体表达与纯化研究. 中国生物工程杂志, 2006, 26(9): 32~37
Qiao J J, Liu H B, Zhu J J, et a1. China Biotechnology, 2006, 26(9): 32~37
［8］ 朱娟娟,荣子烨,江冰,等.抗ErbB2嵌合抗体chA21聚集现象的鉴定和分析. 生物工程学报, 2008, 24(11):1918~1923
Zhu J J, Rong Z Y, Jiang B, et a1. Chinese Journal of Biotechnology, 2008, 24(11):1918~1923
［9］ 王军志. 生物技术药物研究开发和质量控制. 北京: 科学出版社,2002,139~165
Wang Z J. Research& Development and Quality Control of Bio-technical Drug, Beijing:Science Press,2002,139~165
［10］ Hahn R, Bauerhansl P, Shimahara K, et a1. Comparison of protein A affinity sorbents:II. Mass transfer properties. J Chromatogr A, 2005, 1093(1-2): 98~110
［11］ Hahn R, Shimahara K, Steindl F, et a1. Comparison of protein A affinity sorbents:III. Life time study．J Chromatogr A, 2006, 1102(1-2): 224~231
［12］ Wei W. Protein aggregation and its inhibition in biopharmaceutics. Int J Pharm, 2005, 289(1-2): 1~30
［13］ Shukla A A, Gupta P, Han X. Protein aggregation kinetics during Protein A chromatography:Case study for an Fc fusion protein. J Chromatogr A, 2007, 1171(1-2):22~28
［14］ Pete Gagnon. Improved antibody aggregate removal by hydroxyapatite chromatography in the presence of polyethylene glycol. J Immunol Methods, 2008, 336 (2): 222~228
［15］ Wan Y, Vasan S, Ghosh R,et al. Separation of monoclonal antibody alemtuzumab monomer and dimers using ultrafiltration. Biotechnol Bioeng, 2005, 90(4):422~432
［16］ Aldington S,Bonnerjea J. Scale-up of monoclonal antibody purification processes. J Chromatogr B, 2007, 848(1): 64~78
［17］ Baselga J, Norton L,Albanell J, et al. Recombinant humanized anti-HER2 antibody (Herceptin) enhances the antitumor activity of paclitaxel and doxorubicin against HER2/neu overexpressing human breast cancer xenografts. Cancer Res, 1998, 58(13):2825~2831
|
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
