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目的制备人胸苷激酶单克隆抗体。方法用大肠杆菌表达和纯化的人胸苷激酶免疫Balb/C小鼠,免疫小鼠脾细胞与骨髓瘤细胞融合,HAT选择培养和ELISA筛选阳性克隆,阳性克隆杂交瘤注入小鼠腹腔制备腹水,用ELISA和免疫组化鉴定胸苷激酶单克隆抗体。结果经细胞融合和筛选得到37个阳性单克隆杂交瘤,其中有二株杂交瘤可以稳定地产生人胸苷激酶的特异性抗体,制备的腹水中有高效价的抗体活性。结论用原核表达的人胸苷激酶免疫小鼠可以制备出人胸苷激酶特异性单克隆抗体。
Objective To prepare human thymidine kinase monoclonal antibody. Methods Balb / C mice were immunized with human thymidine kinase expressed and purified by Escherichia coli. The spleen cells of immunized mice were fused with myeloma cells. The positive clones were selected by HAT selection and positive clones were screened by ELISA. The positive clonal hybridomas were injected into abdominal cavity of mice to prepare ascites, Identification of thymidine kinase monoclonal antibodies by ELISA and immunohistochemistry. Results 37 positive monoclonal hybridomas were obtained by cell fusion and screening. Among them, two hybridomas could stably produce the specific antibody of human thymidine kinase, and the prepared ascites had high titer antibody activity. Conclusion Human thymidine kinase-specific monoclonal antibodies can be prepared by immunization of mice with human thymidine kinase.