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Logo of bmcbiotBioMed Centralsearchsubmit a manuscriptregisterthis articleBMC Biotechnology
 
BMC Biotechnol. 2012; 12: 70.
Published online Oct 5, 2012. doi:  10.1186/1472-6750-12-70
PMCID: PMC3487952
Enhanced cell disruption strategy in the release of recombinant hepatitis B surface antigen from Pichia pastoris using response surface methodology
Yew Joon Tam,1,2 Zeenathul Nazariah Allaudin,corresponding author1,2 Mohd Azmi Mohd Lila,1 Abdul Rani Bahaman,1 Joo Shun Tan,2 and Morvarid Akhavan Rezaei1,2
1Department of Veterinary Pathology and Microbiology, Faculty of Veterinary Medicine, Universiti Putra Malaysia, Serdang, 43400, Selangor, Malaysia
2Laboratory of Immunotherapeutic and Vaccine Technology (LIVES), Institute of Bioscience, Universiti Putra Malaysia, Serdang, Selangor, Malaysia
corresponding authorCorresponding author.
Yew Joon Tam: yjtam77/at/gmail.com; Zeenathul Nazariah Allaudin: zeenathul/at/putra.upm.edu.my; Mohd Azmi Mohd Lila: azmi/at/vet.upm.edu.my; Abdul Rani Bahaman: rani/at/vet.upm.edu.my; Joo Shun Tan: jooshun/at/gmail.com; Morvarid Akhavan Rezaei: morvaridakhavan/at/yahoo.com
Received March 9, 2012; Accepted October 2, 2012.
Abstract
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.
Keywords: Hepatitis B surface antigen, Cell disruption, Glass bead, High pressure homogenizer, Pichia pastoris, Recombinant protein
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