提供抗类风湿关节炎实验工具:人Janus激酶1单克隆抗体的免疫学特性

Preparation of anti-rheumatoid arthritis experimental instrument: immunological characters of monoclonal antibodies against human Janus kinase 1

目的:制备人Janus激酶1单克隆抗体,并鉴定其免疫学特性。方法:实验于2003-03/2005-04在湖南远泰生物技术有限公司免疫学实验室完成。用化学合成的部分人Janus激酶1多肽链与锁眼蓝蛋白偶联后的全抗原对BALB/c小鼠进行免疫,采用杂交瘤技术制备抗人Janus激酶1的单克隆抗体;同时采用酶联免疫吸附法和免疫印迹法进行筛选;免疫印迹法和免疫细胞化学分析鉴定单克隆抗体的特异性。结果:获得了一株可稳定分泌人Janus激酶1单克隆抗体的细胞株4E5D4,单抗的Ig亚类为IgG。通过免疫印迹法检测,发现其能够特异性识别人Janus激酶1蛋白。结论:成功制备了抗人Janus激酶1的单克隆抗体,为进一步研究其对类风湿关节炎的临床疗效提供了实验工具。

AIM： To prepare monoclonal antibodies （MAbs） against human Janus kinase 1 （bJAK1）, and identify their immunological characters. METHODS： The experiment was carried out in the immunological laboratory of ProMab Biotechnalogical Technology Company between March 2003 and April 2005. BALB/c mice were immunized with the conjugant antigen （the C-terminal peptides of hJAK1 conjugated with KLH）, and then MAbs were prepared by hybridoma technique. Single cell lines which stably produce antibodies against hJAK1 was picked out by enzyme linked immunosorbent assay （ELISA） and western-blot analysis, Western-blot analysis with the antigen hJAK1 and immunocytochemistry （1CC） were used to identify specificity, RESULTS： One hybridoma cell lines named as 4E5D4 secreting hJAK1 MAb was obtained. Its Ig subgroup belonged to lgG. The MAb which could recognize hJAK1 was tested by western-blot and ICC, CONCLUSION： One MAb against hJAK1 had been prepared successfully and possessed high titers, specificity and relative affinity, Preparation of anti-hJAK1 MAb provided a useful reagent for further studying the clinical effect on the new anti-rheumatold arthritis drug.