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目的：制备人精浆蛋白特异性单链抗体,并检测其生物活性与靶向性。方法：应用快速翻译系统(RTS)表达γ-精浆蛋白特异性单链抗体,制备E4B7ScFv肽段和纳米颗粒偶联物,应用免疫沉淀反应、Western Blotting法、免疫荧光染色分析和细胞实验,研究该抗体进入前列腺癌细胞的活性。结果：成功地制备了抗γ-精浆蛋白特异性单链抗体纳米颗粒偶联物。该抗体可与前列腺癌细胞特异性结合,并可在15 min内将纳米磁珠带入前列腺癌细胞内,进入癌细胞的磁球数量随着细胞培养时间的延长而增多,对照实验显示该偶联物不能进入其它组织肿瘤细胞。结论：制备的抗γ-精浆蛋白特异性单链抗体生物活性与靶向性均较好,在前列腺癌的早期核磁共振显像诊断与靶向治疗上具有潜在的应用价值。
Objective: To prepare human serum-specific single-chain antibody and to test its biological activity and targeting. METHODS: E4B7ScFv peptide and nanoparticle conjugate were prepared by using rapid translation system (RTS) to express γ-seminoprotein specific single chain antibody. Immunoprecipitation, Western Blotting, immunofluorescence and cell assays were used to study The antibody enters prostate cancer cells. Results: An anti-γ-seminoprotein specific single chain antibody nanoparticle conjugate was successfully prepared. The antibody specifically binds to prostate cancer cells and can bring nanomagnetic beads into prostate cancer cells within 15 minutes. The number of magnetic balls entering the cancer cells increases with the time of cell culture, and the control experiment shows that the antibody The conjugate can not enter other tissue tumor cells. CONCLUSION: The prepared anti-γ-seminoprotein specific scFv has good biological activity and targeting activity, and it has potential value in diagnosis and targeted therapy of early stage prostate cancer.