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目的:制备具有肿瘤组织特异性表达的L1-ORF1蛋白多克隆抗体并进行初步应用研究。方法:采取基因工程表达方法制备L1-ORF1蛋白,免疫家兔制备多克隆抗体,间接ELISA检测抗体效价,Western blot和细胞免疫荧光方法检测抗体特异性,免疫检测验证其识别肿瘤细胞内L1-ORF1蛋白的特异性。结果:制备的抗L1-ORF1蛋白多克隆抗体具有很高的敏感性与特异性,免疫学检测表明该抗体不仅能检测出正常细胞中瞬时表达的L1-ORF1蛋白,而且可检测出肿瘤细胞中天然表达的L1-ORF1蛋白。结论:制备的多克隆抗体具有较高的敏感性与特异性,为以后该抗体的进一步应用奠定了基础。
OBJECTIVE: To prepare polyclonal antibody of L1-ORF1 protein with tumor-specific expression and to conduct preliminary application research. METHODS: L1-ORF1 protein was prepared by gene engineering method, and polyclonal antibody was prepared by immunization of rabbits. The antibody titer was detected by indirect ELISA. The antibody specificity was detected by Western blot and immunofluorescence assay. ORF1 protein specificity. Results: The anti-L1-ORF1 polyclonal antibody was highly sensitive and specific. Immunological detection showed that the antibody could not only detect the L1-ORF1 protein transiently expressed in normal cells, but also detect tumor cells Naturally expressed L1-ORF1 protein. Conclusion: The prepared polyclonal antibody has high sensitivity and specificity, which lays the foundation for further application of the antibody.