抗人SOCS1单克隆抗体的制备及其生物学特性的鉴定

目的锏备高效价的抗人SOCS1单克隆抗体并对其生物学特性进行鉴定。方法以重组GST-SOCS1融合蛋白免疫BALB／c小鼠，取其脾细胞与Sp2／0细胞融合，经多次筛选及克隆化，建立可稳定分泌抗SOCS1单克隆抗体的杂交瘤细胞株。用ELISA及Westernb1ot鉴定单克隆抗体的特性，并测定其效价、Ig亚类及相对亲和力。结果筛选到一株可稳定分泌抗人SOCS1单克隆抗体的杂交瘤细胞株。Westernb1ot显示在相对分子量为6．0×10^4处出现特异性反应带。杂交瘤细胞培养上清效价为1：1280，腹水效价为1；102400，Ig亚类为IgG1，相对亲和力达1．17×10^7L／mol。结论成功制备出能特异性识别人SOCS1的单克隆抗体，为进一步研究人体细胞因子信号传导的负反馈调节及SOCS1在微生物感染中的免疫调节作用奠定了基础。

Preparation and identification of monoclonal antibody against human SOCS1

Objective To obtain and identify the monoelonal antibody （McAb） against human SOCS1. Methods BALB/c mice were immunized with recombinant GST-SOCS1 protein. Sp2/0 cells were fused with spleen cells of immunized mice. After screened and cloned repeatedly, the strain of hybridoma cell secreting anti-SOCS1 McAb was obtained. The specificity of anti-SOCS1 McAb was checked and evaluated with ELISA and Western blot. Its titer, immunoglobulin subtype and affinity were also measured. Results A strain of hybridoma cell secreting anti-SOCS1 McAb was successfully obtained and identified belong to IgG1 subtype. Its titers in cultural supernatant and ascetic fluid were 1 ： 1280 and 1 ：102400, respectively, by ELISA. Its affinity reached 1.17×10^7 L/mol. Conclusion The anti-SOCS1 McAb, which was successfully prepared, provides the foundation for further study on the negative regulation of cytokine signal transduction and the immuno-regulation of infected diseases caused by microorganisms.