OAS1蛋白单克隆抗体的制备及其表征

目的:用OAS1蛋白免疫小鼠,获得OAS1特异性单克隆抗体,为OAS1的含量测定提供基础。方法:通过全基因合成的方法获得目的基因序列,转化大肠杆菌BL21细胞诱导His-OAS1及OAS1蛋白表达,纯化后用作抗原免疫小鼠,取脾融合,筛选稳定分泌抗体的阳性细胞株,制备并纯化单抗,通过SDS-PAGE,ELISA,Western blot等方法进行检测。结果:体外高效表达OAS1蛋白,并成功制备特异性单克隆抗体,效价在5×10^-11mol/L以上,亲和常数为3.37×10⁸ L·mol^-1。结论:获得高亲和力OAS1单克隆抗体,为其含量的检测奠定了基础。

Production and Characterization of Anti-OAS1 Monoclonal Antibody

Aim: To detect OAS1, its monoclonal antibody was prepared by immune mouse. Methods: BALB/c female mice were immunized with His-OAS1 expressed by E.coli BL21 cells. Monoclonal antibody (McAb) was prepared by hybridoma technology. Identify the purity and characteristics of the selected McAb purified with saturated ammonium sulfate and give preliminary evaluation of the effectiveness of its application, in which SDS-PAGE, ELISA, Western blot and other methods were used. Results: High efficient expression of OAS1 protein in vivo, a McAb can detect the OAS1 was obtained designated as C24-5. The titer of C24-5-McAb was over 9×10^-10mol/L, and the affinity constant of C24-5-McAb was 7.66×10⁷ L·mol^-1. Conclusion: The preparation of C24-5-McAb is very important to further improve the detection of OAS1.