抗人tPA单克隆抗体的制备

PREPARATION AND CHARACTERIZATION OF tPA MONOCLONAL ANTIBODY

目的：制备抗ｔＰＡ单克隆抗体，用于建立ＥＬＩＳＡ方法和研究ｔＰＡ与肿瘤转移的关系。方法：用基因重组的组织型纤溶酶原激活剂（ｒｔＰＡ）免疫Ｂａｌｂ／ｃ小鼠，按常规法作细胞融合，用ＥＬＩＳＡ法筛选出阳性反应的杂交瘤细胞克隆。结果：获得一株杂交瘤细胞株（２Ｂ７）；其免疫球蛋白亚类为ＩｇＧ１；经ＥＬＩＳＡ法和免疫印迹技术证明，此单抗能特异性识别ｔＰＡ。结论：用２Ｂ７ＭｃＡｂ特异性强，用其建立的夹心ＥＬＩＳＡ检测ｔＰＡ的方法线性较好，灵敏度可达１ｎｇ／ｍｌ。２Ｂ７ＭｃＡｂ可望用于ｔＰＡ与肿瘤转移关系及导向溶栓的研究。

Objective:In order to prepare anti-tPA monoclonal antibody,which is used to study the role of tPA in tumor metastasis.Methods:Babl/c was immunized peritoneally with rtPA and complete con-jugate.Then,immunized spleen cells and SP2/0 were fused in the help of PEG,Positive clones were selected with ELISA. Results:One hybridoma(2B7)secreting specific anti-tPA McAb was prepared.The McAb is subclassed into IgG1.Conclusions:2B7 McAb react specifically with both recombinant and natural tPA,but not with irrelevant proteins,ELISA kit established with 2B7 McAb showed high sen-sitivity and specificity.