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目的:研制抗SARSCoVN蛋白的单克隆抗体(mAb)。方法:以纯化的GSTN免疫BALB/c小鼠,采用杂交瘤技术制备抗SARSCoVN蛋白mAb;用免疫双扩散鉴定Ig亚类;Westernblot和免疫组化鉴定mAb的特异性;间接ELISA检测mAb的腹水效价、相对亲和常数。结果:获得1株可分泌特异性mAb的抗SARSCoVN蛋白的杂交瘤细胞系3E10H,Ig亚类为IgG2b;其腹水效价为8×10-5;其相对亲和力1.725×10-10mol/L,Westernblot和免疫组化阳性。结论:获得特异性抗SARSCoVN蛋白的mAb,为进一步用于临床诊断和实验研究创造了条件。
Objective: To develop a monoclonal antibody (mAb) against SARSCoVN protein. Methods: BALB / c mice were immunized with purified GSTN, anti-SARSCoVN mAb was prepared by hybridoma technique, Ig subclass was identified by double immunodiffusion, and the specificity of mAb was identified by Western blot and immunohistochemistry. Price, relative affinity constant. Results: 1 anti-SARSCoVN protein-secreting mAb 3E10H was secreted and the Ig subclass was IgG2b. The ascites titer was 8 × 10-5. The relative affinity was 1.725 × 10-10mol / L. Western blot And immunohistochemistry positive. Conclusion: Obtaining mAbs specific for anti-SARSCoVN protein can be used for further clinical diagnosis and experimental research.