国家科技攻关计划项目(中国人类肝脏蛋白质组计划,863计划,973项目,2004BA711A20).
This work was supported by a grant from The National Science and Technology Research and Development Program of China(2004BA711A20)
以表面展示人源蛋白酶体α亚基6蛋白的重组酵母细胞,结合酶联吸附免疫原理检测技术,以表面展示有人源蛋白酶体α亚基6 (proteasome subunit alpha 6,PSα 6) 的重组酵母细胞及其对应的单克隆抗体3D7D12D为研究对象,建立酵母酶联免疫吸附 (yeast-ELISA) 检测技术,应用于检测小鼠单克隆抗体及单抗效价. 应用该方法最佳酵母浓度为0.50 A600;测得单抗效价为1∶5×105,与常规ELISA效价接近;交叉试验和阻断试验表明该方法特异性强;同时该方法可用于检测抗血清.结果表明,yeast-ELISA可直接应用于检测单抗,测得效价与常规抗原包被间接ELISA具有良好一致性,特异性好,无交叉反应性.
With Saccharomyces cerevisiae cell surface engineering, combining with enzyme-linked immunosorbent assay (ELISA), a new method has been, which is named as ‘yeast-ELISA’ detection. Here, the recombination yeast cell MT8-1 surface display expressing antigen HPSα6 (Human proteasome subunit alpha 6), and its monoclonal antibody 3D7D12D4 (hybridoma cell line No) was used as study object. The fitful yeast cell concentration of this method is 0.50 A600, and it was used to detect monoclonal antibody and its valance (1∶5×105). The specificity and sensitivity of the method was demonstrated by crossing test and interdiction test. The results indicated that ‘yeast-ELISA’ could detect monoclonal antibody directly, and the valance of which is concordance with the result of the proteantigen indirect ELISA. And also this method could be used to antiserum detection, which also has good result. It demonstrates that the method of the ‘yeast-ELISA’ detection was established, which is the supply and application of Saccharomyces cerevisiae cell surface engineering.
叶茂,韩双艳,林影,唐语谦,王小宁.酵母表面展示-酶联免疫吸附测定法的建立[J].生物化学与生物物理进展,2007,34(10):1113-1117
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