Enhanced cell disruption strategy in the release of recombinant hepatitis B surface antigen from Pichia pastoris using response surface methodology

BMC Biotechnology

Research Article

Mohd Azmi Mohd Lila¹  Abdul Rani Bahaman¹  Yew Joon Tam²  Zeenathul Nazariah Allaudin²  Morvarid Akhavan Rezaei²  Joo Shun Tan³ 

[1] Department of Veterinary Pathology and Microbiology, Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400, Serdang, Selangor, Malaysia;Department of Veterinary Pathology and Microbiology, Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400, Serdang, Selangor, Malaysia;Laboratory of Immunotherapeutic and Vaccine Technology (LIVES), Institute of Bioscience, Universiti Putra Malaysia, Serdang, Selangor, Malaysia;Laboratory of Immunotherapeutic and Vaccine Technology (LIVES), Institute of Bioscience, Universiti Putra Malaysia, Serdang, Selangor, Malaysia;

关键词: Hepatitis B surface antigen;    Cell disruption;    Glass bead;    High pressure homogenizer;    Pichia pastoris;    Recombinant protein;   

DOI  :  10.1186/1472-6750-12-70

 received in 2012-03-09, accepted in 2012-10-02,  发布年份 2012

来源: Springer

【 摘 要 】

BackgroundCell disruption strategies by high pressure homogenizer for the release of recombinant Hepatitis B surface antigen (HBsAg) from Pichia pastoris expression cells were optimized using response surface methodology (RSM) based on the central composite design (CCD). The factors studied include number of passes, biomass concentration and pulse pressure. Polynomial models were used to correlate the above mentioned factors to project the cell disruption capability and specific protein release of HBsAg from P. pastoris cells.ResultsThe proposed cell disruption strategy consisted of a number of passes set at 20 times, biomass concentration of 7.70 g/L of dry cell weight (DCW) and pulse pressure at 1,029 bar. The optimized cell disruption strategy was shown to increase cell disruption efficiency by 2-fold and 4-fold for specific protein release of HBsAg when compared to glass bead method yielding 75.68% cell disruption rate (CDR) and HBsAg concentration of 29.20 mg/L respectively.ConclusionsThe model equation generated from RSM on cell disruption of P. pastoris was found adequate to determine the significant factors and its interactions among the process variables and the optimum conditions in releasing HBsAg when validated against a glass bead cell disruption method. The findings from the study can open up a promising strategy for better recovery of HBsAg recombinant protein during downstream processing.

【 授权许可】

Unknown   
© Tam et al.; licensee BioMed Central Ltd. 2012. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.