Some factors influencing the proportion of periplasmic hepatitisB virus pre-S2 antigen in the recombinant yeastHansenula polymorpha |
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Authors: | Marie-Renée de Roubin Michael D Cailas Shi-Hsiang Shen Denis Groleau |
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Institution: | (1) Biotechnology Research Institute, National Research Council of Canada, 6100 Royalmount Avenue, H4P 2R2 Montreal, Quebec, Canada;(2) Environmental and Occupational Health Sciences, University of Illinois, School of Public Health, Chicago, Illinois, U.S.A. |
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Abstract: | Summary A central composite design (CCD) was used to evaluate, for the purpose of future process optimization, the influence of pH, yeast extract and ammonium chloride concentrations on the proportion of periplasmic hepatitisB pre-S2 antigen in the recombinant yeastHansenula polymorpha. Each factor was tested at five levels, and a second order polynomial model for the proportion of periplasmic antigen was fitted to the treatment combinations. pH showed the greatest effect: the proportion of periplasmic antigen was greatly increased at the higher pH levels. At the higher pH levels used, the proportion of periplasmic antigen was enhanced by a high concentration of ammonium chloride. Additional experiments have confirmed both the validity of the selected model and the optimal conditions found. A significant correlation was found between the proportion of periplasmic antigen and the total yield of antigen. These results indicated that is should be possible to modulate the distribution of the pre-S2 antigen between the periplasm and the cytoplasm of the yeast. |
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Keywords: | Secretion Periplasmic pre-S2 antigen Recombinant protein Experimental design Methylotrophic yeast |
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