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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
2007-12-01
Source PublicationPROTEIN EXPRESSION AND PURIFICATION
ISSN1046-5928
Volume56Issue:2Pages:301-310
Abstract

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.

KeywordPurification Recombinant Hepatitis b Surface Antigen Hansenula Polymorpha Integrated Chromatographic Process
SubtypeArticle
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
DOI10.1016/j.pep.2007.08.009
Indexed BySCI
Language英语
WOS KeywordChinese-hamster Ovary ; Size-exclusion Chromatography ; Ion-exchange Chromatography ; Laser-light Scattering ; Cell-line ; Expression ; Immunogenicity ; Aggregation ; Particles ; Vector
WOS Research AreaBiochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
WOS SubjectBiochemical Research Methods ; Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
WOS IDWOS:000251312300020
Citation statistics
Document Type期刊论文
Identifierhttp://ir.ipe.ac.cn/handle/122111/19217
Collection生化工程国家重点实验室
Affiliation1.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
Recommended Citation
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.
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