Comparative evaluation of different cell disruption methods for the release of recombinant hepatitis B core antigen from <Emphasis Type="Italic">Escherichia coli</Emphasis> |
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Authors: | Chin Woi Ho Wen Siang Tan Wei Boon Yap Tau Chuan Ling Beng Ti Tey |
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Institution: | (1) Division of Autotrophic Microorganisms, Institute of Microbiology, Academy of Sciences of the Czech Republic, 379 81 T?eboň, Czech Republic |
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Abstract: | A comparative evaluation of five different cell-disruption methods for the release of recombinant hepatitis B core antigen
(HBcAg) from Escherichia coli was investigated. The cell disruption techniques evaluated in this study were high-pressure homogenization, batch-mode bead
milling, continuous-recycling bead milling, ultrasonication, and enzymatic lysis. Continuous-recycling bead milling was found
to be the most effective method in terms of operating cost and time. However, the highest degree of cell disruption and amounts
of HBcAg were obtained from the high-pressure homogenization process. The direct purification of HBcAg from the unclarified
cell disruptate derived from high-pressure homogenization and bead milling techniques, using batch anion-exchange adsorption
methods, showed that the conditions of cell disruption have a substantial effect on subsequent protein recovery steps. |
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Keywords: | hepatitis B core antigen high-pressure homogenization bead milling ultrasonication lysozyme batch anion-exchange adsorption |
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