A highly efficient integrated chromatographic procedure for the purification of recombinant hepatitis B surface antigen from Hansenula polymorpha
文献类型：期刊论文
| 作者 | Huang, Yongdong; Bi, Jingxiu; Zhang, Yan; Zhou, Weibin; Li, Yan; Zhao, Lan; Su, Zhiguo |
| 刊名 | PROTEIN EXPRESSION AND PURIFICATION
 |
| 出版日期 | 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 |
| 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. |
入库方式： OAI收割
来源：过程工程研究所
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