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Improving recovery of recombinant hepatitis B virus surface antigen by ion-exchange chromatographic supports with low ligand density

Paper ID Volume ID Publish Year Pages File Format Full-Text
36115 45121 2006 7 PDF Available
Title
Improving recovery of recombinant hepatitis B virus surface antigen by ion-exchange chromatographic supports with low ligand density
Abstract

Ion-exchange chromatography (IEC) with the lowest step recovery (<50%) is still a bottleneck of the essentially four-step chromatographic procedure for the purification of recombinant hepatitis B surface antigen (r-HBsAg) from transformed Chinese hamster ovary (CHO) cell line. The low recovery is mainly resulted from the change of the assemble structure of HBsAg particles such as disassociation or aggregation on the surface of the absorbent during binding and eluting procedures. To improve the chromatographic efficiency of IEC, the influence of ligand density from 0.041 to 0.130 mmol diethylaminoethyl (DEAE)/ml absorbent on IEC of r-HBsAg was investigated. The retention time of HBsAg particles depended highly on ligand density. Higher HBsAg recovery and purification factor (PF) were obtained from the absorbents with lower ligand density. Under the optimum chromatographic conditions, the HBsAg recovery was increased from about 44% to 66% and the PF was increased from 9 to 13 by decreasing the ligand density from 0.130 to 0.041 mmol DEAE/ml absorbent.

Keywords
Recombinant hepatitis B surface antigen; Ion-exchange chromatography; Ligand density; Multi-point interaction
First Page Preview
Improving recovery of recombinant hepatitis B virus surface antigen by ion-exchange chromatographic supports with low ligand density
Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 41, Issue 11, November 2006, Pages 2320–2326
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering