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A highly efficient hydrophobic interaction chromatographic absorbent improved the purification of hepatitis B surface antigen (HBsAg) derived from Hansenula polymorpha cell

Paper ID Volume ID Publish Year Pages File Format Full-Text
35976 45117 2007 6 PDF Available
Title
A highly efficient hydrophobic interaction chromatographic absorbent improved the purification of hepatitis B surface antigen (HBsAg) derived from Hansenula polymorpha cell
Abstract

To improve the purification efficiency of recombinant hepatitis B surface antigen derived from Hansenula polymorpha (Hans-HBsAg), a serial of absorbents for hydrophobic interaction chromatography with the controllable ligand density and spacer arm were synthesized, then developed and further applied to purify Hans-HBsAg. The absorbent, 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 absorbent 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 chromatographic 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; Hydrophobic interaction chromatography; Hansenula polymorpha; Ligand density; Absorbent
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A highly efficient hydrophobic interaction chromatographic absorbent improved the purification of hepatitis B surface antigen (HBsAg) derived from Hansenula polymorpha cell
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Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 42, Issue 5, May 2007, Pages 751–756
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us