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目的 制备针对结直肠癌缺陷 ( deleted colorectal carcinom a,DCC)蛋白抗原决定簇多肽的单克隆抗体。方法 用合成的 DCC蛋白抗原决定簇多肽 (氨基酸序列从 195～ 2 0 4)为抗原 ,免疫 BAL B/c小鼠 ,经细胞融合 ,有限稀释法筛选出能分泌抗 DCC合成肽单克隆抗体的杂交瘤细胞株 ,并对单克隆抗体进行了初步鉴定。结果 细胞融合率 75 % ,经 3次亚克隆 ,最后保留一株能稳定分泌抗 DCC合成肽单克隆抗体的杂交瘤细胞株 ( NJ99)。免疫印迹及组织定位显示该单克隆抗体特异性高、亲和力强。结论 抗 DCC合成肽单克隆抗体杂交瘤细胞株的建立为深入研究 DCC表达及临床应用提供了有力的工具
Objective To prepare monoclonal antibodies against epitopes of deleted colorectal carcinom a (DCC) protein. Methods BALB / c mice were immunized with a synthetic DCC protein epitope peptide (amino acid sequence from 195 to 204). The monoclonal antibodies secreting anti-DCC peptide were screened by limiting dilution method Hybridoma cell lines, and monoclonal antibodies were initially identified. Results The rate of cell fusion was 75%. After three times of subcloning, a hybridoma cell line (NJ99) which can stably secrete monoclonal antibody against DCC synthetic peptide was reserved. Western blotting and tissue localization showed that the monoclonal antibody has high specificity and strong affinity. Conclusion The establishment of monoclonal antibody against DCC synthetic peptide hybridoma cell lines for the study of DCC expression and clinical application provides a powerful tool