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Efficient mechanical cell disruption of Escherichia coli by an ultrasonicator and recovery of intracellular hepatitis B core antigen

Paper ID Volume ID Publish Year Pages File Format Full-Text
36206 45124 2006 6 PDF Available
Title
Efficient mechanical cell disruption of Escherichia coli by an ultrasonicator and recovery of intracellular hepatitis B core antigen
Abstract

Hepatitis B core antigen (HBcAg) expressed intracellularly in Escherichia coli, hence cell disruption process is the prerequisite step to recover this protein. The objective of this study was to develop an efficient mechanical cell disruption method (ultrasonication) for the release of HBcAg. The released intracellular protein was subsequently purified by the combination of centrifugation, precipitation, dialysis and sucrose gradient ultracentrifugation. The results show that the cell disruption and HBcAg release rates increased with the increase of acoustic power. Besides, the optimal cell suspension volume (15 ml) and sonication time (90 min) for the disruption of E. coli cells and the release of intracellular HBcAg were determined. Ultrasonication was found to be more effective than enzymatic method with respect to the release of HBcAg. HBcAg recovered from the cell lysate, resulted from ultrasonication, is 22 times higher than that of the enzymatic method. Electron microscopic analysis showed that the purified HBcAg assembled into particles that closely resemble the viral nucleocapsid form. These particles reacted with anti-HBcAg monoclonal antibody in enzyme-linked immunosorbent assay (ELISA) showing that it is functionally active.

Keywords
Cell disruption; HBcAg; Ultrasonication; Lysozyme; Escherichia coli
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Efficient mechanical cell disruption of Escherichia coli by an ultrasonicator and recovery of intracellular hepatitis B core antigen
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Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 41, Issue 8, August 2006, Pages 1829–1834
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