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目的:应用眼镜王蛇毒基因疫苗,免疫BALB/c小鼠,制备单克隆抗体(mAb)。方法:应用眼镜王蛇Oh-3基因真核表达质粒pIRES-Oh3,佐以脂质体制成基因疫苗,免疫BALB/c小鼠,取其脾细胞与同源Sp2/0骨髓瘤细胞进行融合,经间接ELISA筛选,有限稀释法克隆,制备mAb。结果:获得了2株分泌抗眼镜王蛇毒mAb的杂交瘤细胞株(F5、F11),细胞培养上清液的抗体效价分别为3.2×10-1、6.4×10-1,腹水抗体效价分别为1.28×10-4、2.56×10-4。结论:成功地制备了2株眼镜王蛇毒mAb。
OBJECTIVE: Monoclonal antibody (mAb) was prepared by immunizing BALB / c mice with the king cobra venom gene vaccine. Methods: The eukaryotic expression plasmid pIRES-Oh3 of Oh-3 gene was co-cultured with liposome to immunize BALB / c mice. The spleen cells were fused with sp2 / 0 myeloma cells. After indirect ELISA screening, limiting dilution cloning, preparation of mAb. Results: Two hybridoma cell lines secreting McAb against King Cobra were obtained (F5 and F11). The antibody titers of the cell culture supernatant were 3.2 × 10-1 and 6.4 × 10-1, respectively. The antibody titer Respectively 1.28 × 10-4 and 2.56 × 10-4. Conclusion: Two strains of cobra venom mAb were successfully prepared.