title: Enhanced cell disruption strategy in the release of recombinant hepatitis B surface antigen from Pichia pastoris using response surface methodology.
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.
authors.affiliation: Department of Veterinary Pathology and Microbiology, Faculty of Veterinary Medicine, Universiti Putra Malaysia, Serdang, 43400, Selangor, Malaysia.
authors.forename: Yew Joon
authors.initials: YJ
authors.lastname: Tam
authors.forename: Zeenathul Nazariah
authors.initials: ZN
authors.lastname: Allaudin
authors.forename: Mohd Azmi Mohd
authors.initials: MA
authors.lastname: Lila
authors.forename: Abdul Rani
authors.initials: AR
authors.lastname: Bahaman
authors.forename: Joo Shun
authors.initials: JS
authors.lastname: Tan
authors.forename: Morvarid Akhavan
authors.initials: MA
authors.lastname: Rezaei
mesh_terms: D001696:Biomechanical Phenomena; D001703:Biophysics / Q000379:methods*; D015870:Gene Expression; D006514:Hepatitis B Surface Antigens / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D007218:Industrial Microbiology / Q000379:methods*; D010843:Pichia / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D011312:Pressure; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D006514:Hepatitis B Surface Antigens; D011994:Recombinant Proteins