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作者用基因重组方法,将含有乙型肝炎病毒(HBV)S基因的质粒载体连接到酵母乙醇脱氢酶Ⅰ启动子上,通过质粒的自身复制,在酿酒酵母表面表达出一种非糖基化的多肽p25。它具有完全免疫原性,能诱生抗-HBs应答。为了稳定地保持p25多肽的抗原活性,作者进一步研制具有高度免疫原性的多肽微胶粒。将纯化的酵母HBsAg颗粒悬液高速离心,并重新悬浮于pH7.5 0.01M Tris缓冲液内,使最终浓度达1mg/ml。用非离子去垢剂Triton X-100破坏HBsAg的脂质双层,然后在20～60%蔗糖梯度中分层,制成多肽微胶
The authors used gene recombination method, the hepatitis B virus (HBV) S gene plasmid vector is connected to the yeast alcohol dehydrogenase I promoter, through the plasmid self-replication, in Saccharomyces cerevisiae surface expression of a non-glycosylation Of the polypeptide p25. It is completely immunogenic and can induce anti-HBs responses. In order to stably maintain the antigenic activity of the p25 polypeptide, the authors further developed highly immunogenic peptide micelles. The purified yeast HBsAg particle suspension was centrifuged at high speed and resuspended in 0.01 M Tris buffer pH 7.5 to a final concentration of 1 mg / ml. The lipid bilayer of HBsAg is disrupted with the nonionic detergent Triton X-100 and then layered in a 20-60% sucrose gradient to make the polypeptide micelles