抗人SOCS1单克隆抗体的制备及其生物学特性的鉴定  

Preparation and identification of monoclonal antibody against human SOCS1

作　　者：陈长春[1,2] 李婉宜[1] 康熙元 姚锋[1] 邝玉[1] 丰锋[1] 

机构地区：[1]四川大学华西基础医学与法医学院微生物学教研室,四川成都610041 [2]广元职工医学院微生物学教研室,四川广元628017

出　　处：《西部医学》2009年第7期1085-1087,1090,共4页Medical Journal of West China

摘　　要：目的制备高效价的抗人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Western blot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。Western blot显示在相对分子量为6.0×104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1.17×107L/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。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.

关 键 词：SOCS1 单克隆抗体 制备及鉴定 杂交瘤细胞 

分 类 号：R392.11[医药卫生—免疫学]