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Many studies have suggested that hepatitis B surface antigen (HBsAg) including PreS sequences could be an ideal candidate for highly effective hepatitis B virus vaccine. Modified surface antigens S1S, SS1 and S2S which carried PreS epitopes, were expressed in Pichia pastoris. The characterization of antigenicity and particle assembly demonstrated that the expression products could be assembled into particles which presented S, PreS1 or PreS2 antigenicity, respectively. The expression was more efficient than that in Saccharomyces cerevisiae.