应用合成多肽技术制备肺癌转移相关蛋白1多克隆抗体的研究

摘要

目的 应用人工合成多肽技术制备肺癌转移相关蛋白1(Lung Cancer Metastasis-Related Protein 1,LCMR1)的多克隆抗体.方法 运用生物信息学技术和人工合成多肽方法制备LCMR1特异性多肽,鉴定纯度后与牛血清白蛋白、匙孔血蓝蛋白进行偶联制备多肽抗原,然后免疫新西兰雄性白兔,获得抗血清,经亲和层析法纯化后,用ELISA法鉴定抗体效价,用Western Blot鉴定特异性.结果 成功获得了LCMR1的多克隆抗体,ELISA检测抗体效价大于1:100000,Western Blot免疫印迹结果显示其具有抗原特异性识别.结论 应用免疫原性肽抗体技术成功制备了LCMR1的多克隆抗体,为今后深入了解LCMR1基因的生物学功能提供了有用的研究工具.%Objective To prepare the polyclonal antibody to lung cancer metastasis-related protein 1 with synthesized peptide. Methods Specific polypeptide of lung cancer metastasis-related protein 1 (LCMR1) was pre-pared by bioinformatics and polypeptide synthesis method, and it was determined by high-performance liquid chroma-tography. Then the polypeptide coupled with bearer protein ( Keyhole Limpet Hemocyanin and Bovine Serum Albu-min) was used to immunize New Zealand Rabbits to prepare serum with anti-LCMR1 polyclonal antibody. The anti-body was purified, and the titer was determined by ELISA and the specificity was identified by Western blot. Results Polyclonal antibody to LCMR1 was obtained successfully, the titer of which was more than 1:100000. Western blot results showed that the antibody specifically recognized LCMR1. Conclusion It is concluded that the polyclonal an-tibody to LCMR1 with high titer and specificity is successfully prepared. The antibody provides an useful experimental tool to investigate the biological function of LCMR1 in future.