国家自然科学基金资助(39400159,39870787)和国家“九五”攻关课题资助项目(96C02-01-17).
克隆分泌CEA杂交瘤细胞重链可变区(VH)和轻链可变区(VL),以Linker连接VH及VL构建抗CEA单链抗体.同时以Spacer连接单链抗体和链亲和素,构建成功单链抗体和链亲和素融合基因,克隆该融合基因至原核表达载体,pET21a(+),经IPTG诱导表达出该双特异性融合蛋白.活性鉴定表明该融合蛋白具有结合CEA及生物素的双特异性.该融合蛋白在生物领域中有较广阔的应用前景.
The expression vector, pET21-CEA-streptavidin was constructed which contains the gene of single chain antibody of CEA(ScFv) and the streptavidin. Constructed vector was expressed by Escherichia coli. The results showed that such protein was successfully prepared and its bispecific activities were testified by affinity blot and Western blot. The biotin binding and immunocytochemistry assay indicated that the expressed fusion protein was capable of binding biotin molecule and CEA antigen respectively. This protein has the potential application when combined with the corresponding tests.
任军,季万胜,周绍娟,陈峥,张学庸,樊代明.抗CEA单链抗体与链亲和素融合基因的表达[J].生物化学与生物物理进展,2000,27(4):390-394
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