A highly efficient integrated chromatographic procedure for the purification of recombinant hepatitis B surface antigen from Hansenula polymorpha

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	A highly efficient integrated chromatographic procedure for the purification of recombinant hepatitis B surface antigen from Hansenula polymorpha
	Huang, Yongdong ; Bi, Jingxiu ; Zhang, Yan ; Zhou, Weibin ; Li, Yan ; Zhao, Lan ; Su, Zhiguo
刊名	PROTEIN EXPRESSION AND PURIFICATION
	2007-12-01
卷号	56 期号:2 页码:301-310
关键词	purification recombinant hepatitis B surface antigen Hansenula polymorpha integrated chromatographic process
ISSN号	1046-5928
英文摘要	The high expression level of recombinant hepatitis B surface antigen obtained from Hansenula polymorpha yeast cell (Hans-HBsAg) made it possible to produce HBsAg vaccine in a large scale and by cost-effective process. However, the present available purification process was somewhat tedious, time-consuming and difficult to scale up. To improve the purification efficiency and simplify the purification process, an integrated chromatographic process was developed and optimized. The downstream process included ion-exchange chromatography (IEC), hydrophobic interaction chromatography (HIC) and gel filtration chromatography (GFC). A series of chromatographic adsorbents were evaluated for their performances on the purification of Hans-HBsAg, and then the suitable adsorbents for IEC and HIC were screened out, respectively. After clarification by centrifugation, the supernatant of cell disruption (SCD) was purified by standard chromatographic steps, IEC on DEAE Sepharose FF, HIC on Butyl-S-QZT and GFC on Sepharose 4FF. Furthermore, HBsAg recovery, purification factor (PF) and purity during the downstream process were evaluated with enzyme-linked immunosorption assay (ELISA), sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and high-performance size-exclusion chromatography (HPSEC). The results demonstrated that in the scale of 550 ml SCD, the total HBsAg recovery and PF of the whole procedure were about 21.0 +/- 0.9% and 80.7 +/- 8.4. (n = 3) respectively, with the purity of above 99%. This new downstream process was efficient, reproducible and relatively easy to be scaled up. (C) 2007 Elsevier Inc. All rights reserved.
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
类目[WOS]	Biochemical Research Methods ; Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
研究领域[WOS]	Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
关键词[WOS]	CHINESE-HAMSTER OVARY ; SIZE-EXCLUSION CHROMATOGRAPHY ; ION-EXCHANGE CHROMATOGRAPHY ; LASER-LIGHT SCATTERING ; CELL-LINE ; EXPRESSION ; IMMUNOGENICITY ; AGGREGATION ; PARTICLES ; VECTOR
收录类别	SCI
语种	英语
WOS记录号	WOS:000251312300020
公开日期	2015-09-01
内容类型	期刊论文
源URL	[http://ir.ipe.ac.cn/handle/122111/19217]
专题	过程工程研究所_生化工程国家重点实验室
作者单位	1.Chinese Acad Sci, Proc Engn Inst, Natl Key Lab Biochem Engn, Beijing 100080, Peoples R China 2.Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China 3.Univ Adelaide, Sch Pharmaceut Engn, Adelaide, SA 5005, Australia
推荐引用方式 GB/T 7714	Huang, Yongdong,Bi, Jingxiu,Zhang, Yan,et al. A highly efficient integrated chromatographic procedure for the purification of recombinant hepatitis B surface antigen from Hansenula polymorpha[J]. PROTEIN EXPRESSION AND PURIFICATION,2007,56(2):301-310.
APA	Huang, Yongdong.,Bi, Jingxiu.,Zhang, Yan.,Zhou, Weibin.,Li, Yan.,...&Su, Zhiguo.(2007).A highly efficient integrated chromatographic procedure for the purification of recombinant hepatitis B surface antigen from Hansenula polymorpha.PROTEIN EXPRESSION AND PURIFICATION,56(2),301-310.
MLA	Huang, Yongdong,et al."A highly efficient integrated chromatographic procedure for the purification of recombinant hepatitis B surface antigen from Hansenula polymorpha".PROTEIN EXPRESSION AND PURIFICATION 56.2(2007):301-310.