摘要
目的制备鼠源性GDDR单克隆抗体,并对其特异性进行鉴定。方法利用SEAL对GDDR氨基酸序列表位行参数评分,结合人、鼠蛋白序列同源性,设计并合成重组GDDR蛋白,并以所获得蛋白免疫BALB/c小鼠制备鼠单克隆抗体,运用ELISA法检测抗体效价,免疫印迹分析和免疫组织化学鉴定抗体特异性。结果经常规细胞融合和筛选获得两株稳定分泌抗GDDR抗体骨髓瘤细胞株,ELISA检测腹水效价达1100000,Western-blot证实抗体可以和GDDR蛋白特异性结合,免疫组织化学结果显示正常胃黏膜组织GDDR表达明显高于胃癌组织。结论成功制备出两株抗GDDR单克隆抗体mAb,为进一步研究GDDR的生物学功能提供良好的工具。
Objective To prepare and identify the monoclonal antibody against murine GDDR protein.Methods The structure of GDDR amino acids was analyzed by SEAL to select the epitope.Recombinant murine GDDR protein was synthesized for immunization of BALB/c mice.The efficiency and specificity of the antibodies were identified by ELISA,Western blot assay and Immunohistochemistry,respectively.Results Two hybridoma cell lines which could secrete antibody stably against GDDR were established.The ascite titers of this monoclonal antibody reached 1100 000,determined by indirect ELISA.The specificity of mAb against GDDR was assessed by Western blot.Immunohistochemistry results showed a higher expression of GDDR in normal gastric tissue than in gastric cancer tissue.Conclusion Two novel murine monoclonal antibodies against GDDR were harvested,and they could be utilized as useful reagents for further study of the biological function of GDDR.
出处
《中华普外科手术学杂志（电子版）》
2012年第4期43-46,共4页
Chinese Journal of Operative Procedures of General Surgery(Electronic Edition)
基金
国家自然科学基金资助项目(81071690)
关键词
基因
肿瘤抑制
序列分析
蛋白质
抗体
单克隆
杂交瘤
Genes
tumor suppressor
Sequenca analysis
protein
Antibodies monoclonal
Hybridomas
