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

BMC Biotechnology

Yew Joon Tam¹  Zeenathul Nazariah Allaudin¹  Mohd Azmi Mohd Lila²  Abdul Rani Bahaman²  Joo Shun Tan¹  Morvarid Akhavan Rezaei¹ 

[1] Laboratory of Immunotherapeutic and Vaccine Technology (LIVES), Institute of Bioscience, Universiti Putra Malaysia, Serdang, Selangor, Malaysia

[2] Department of Veterinary Pathology and Microbiology, Faculty of Veterinary Medicine, Universiti Putra Malaysia, Serdang, 43400, Selangor, Malaysia

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

Others  :  1134764
DOI  :  10.1186/1472-6750-12-70

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

【 摘 要 】

Background

Cell 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.

Results

The 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.

Conclusions

The 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】

   
2012 Tam et al.; licensee BioMed Central Ltd.

【 图 表 】

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