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https://hdl.handle.net/2440/46440
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Type: | Journal article |
Title: | A highly efficient hydrophobic interaction chromatographic absorbentnext term improved the purification of hepatitis B surface antigen (HBsAg) derived from Hansenula polymorpha cell |
Author: | Zhou, W. Bi, J. Zhao, L. Wang, Y. Li, Y. Huang, Y. Ma, G. Su, Z. |
Citation: | Process Biochemistry, 2007; 42(5):751-756 |
Publisher: | Elsevier Sci Ltd |
Issue Date: | 2007 |
ISSN: | 1359-5113 1873-3298 |
Statement of Responsibility: | Weibin Zhou, Jingxiu Bi, Lan Zhao, Yangmu Wang, Yan Li, Yongdong Huang, Guanghui Ma and Zhiguo Su |
Abstract: | To improve the purification efficiency of recombinant hepatitis B surface antigen derived from Hansenula polymorpha (Hans-HBsAg), a serial of previous termabsorbentsnext term for previous termhydrophobic interactionnext term chromatography with the controllable ligand density and spacer arm were synthesized, then developed and further applied to purify Hans-HBsAg. The previous termabsorbent,next term Butyl-S QZT with the ligand density of 25 μmol/(g wet gel) and spacer arm of 3C, was screened out and its physical and chemical properties were evaluated. High rigidity and low backpressure (<0.06 MPa) were obtained at the flow rate up to 20 ml/min. Moreover, it has the stable chemical characteristics of subjecting to high concentrations of acid, alkali and detergents. This HIC previous termabsorbentnext term was further applied to purify Hans-HBsAg with the recovery 94% and purification-fold 9 under the optimized operation condition at pH 6.5 and concentration of ammonium sulfate 7.5%. Finally, the HIC adsorbent of Butyl-S QZT was applied in the integrated three-step previous termchromatographicnext term purification process to purify Hans-HBsAg. About 140 mg of purified Hans-HBsAg was obtained from 1 l cell disruption supernatant at the total recovery of 27% and the purification-fold of 151.8. Based on the assay of SDS-PAGE and SEC-HPLC, the purity of the purified HBsAg was over 99% to meet the requirement for the further inoculation use. |
Keywords: | Recombinant HBsAg Purification previous termHydrophobic interactionnext term chromatography Hansenula polymorpha Ligand density previous termAbsorbentnext term |
DOI: | 10.1016/j.procbio.2007.01.003 |
Description (link): | http://www.elsevier.com/wps/find/journaldescription.cws_home/422857/description#description |
Published version: | http://dx.doi.org/10.1016/j.procbio.2007.01.003 |
Appears in Collections: | Aurora harvest 6 Chemical Engineering publications |
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