肝肠钙粘连蛋白单克隆抗体的制备及其对HepG2的生长抑制作用

目的 制备及鉴定肝肠钙粘连蛋白(CDH17)的单克隆抗体,研究其对肝癌细胞系HepG2的生长抑制作用.方法 利用重组CDH17蛋白为免疫原,免疫BALB/C小鼠,取免疫小鼠的脾细胞和同系小鼠的骨髓瘤细胞Sp2/0进行常规融合,通过间接ELISA的筛选和有限稀释克隆化,获得稳定分泌抗CDH17的单克隆抗体杂交瘤细胞株,通过ELISA和Western blotting分别对其效价和特异性进行鉴定,以不同浓度的单抗分别作用于HepG2细胞24、48、72和96 h.观察细胞数量和形态的变化,MTT法检测其对HepG2细胞的增值抑制作用.结果 成功建立了2株稳定分泌抗CDH17的单克隆抗体杂交瘤细胞株.两株单抗通过Western blotting实验都能特异性地结合真核细胞内源性的CDH17蛋白.不同浓度的单抗作用后细胞数量明显减少、形态不规则.不同浓度的单抗均能抑制HepG2细胞增殖,呈剂量-时间依赖效应,药物浓度和用药时间之间有交互效应(P<0.05).结论 成功建立了两株效价高、特异性好的抗CDH17蛋白的单克隆抗体,其能抑制HepG2细胞增殖,并呈剂量-时间依赖效应.

Effect of monoclonal antibodies against LI-cadherin on the proliferation of human hepatocellular carcinoma cells

Objective To obtain monoclonal antibodies (mAb) against LI-cadherin and investigate their effects on the proliferation of human hepatocellular carcinoma cells. Methods Balb/c mice were immunized with recombinant LI-cadherin,and hybridoma cell lines secreting monoclonal antibodies against LI-cadherin were established with routine cell fusion and subcloning approach. The specificity of these mAbs was determined by enzyme-linked immunosorbent assay (ELISA) and Western blotting. The effect of the mAbs obtained ...