| 目的：制备可特异性识别人晶状体上皮细胞的单克隆抗体，筛选出特异性强亲和力高的克隆。
方法：以原代培养人晶状体上皮细胞经脾内注射免疫BALB/c小鼠，取其脾细胞与NS-1骨髓瘤细胞融合，用间接免疫荧光法以人晶状体上皮细胞为抗原筛选阳性克隆。用免疫荧光、免疫组化法对其特异性进行鉴定。小鼠腹腔接种杂交瘤细胞制备腹水，采用羟基磷灰石色谱法进行纯化。
结果：获得了1株稳定分泌抗人晶状体上皮细胞的杂交瘤细胞系，其亚类为IgM。免疫荧光和免疫组化显示，此抗体与人晶状体上皮细胞膜呈阳性反应，与人眼其他组织呈阴性反应。腹水经羟基磷灰石色谱法纯化，SDS-PAGE检测显示抗体蛋白的纯度为90%。
结论：所获抗体可特异性识别人晶状体上皮细胞，有进一步制备免疫导向药物，用于后囊混浊治疗的应用潜力。
关键词：晶状体上皮细胞；单克隆抗体；杂交瘤细胞；后囊混浊
Objective: To prepare and identify high-affinity monoclonal antibody (McAb) specific to human lens epithelial cell, and preliminarily study its characteristics.
Methods: BALB/c mice were immunized by intrasplenic injection of cultured human lens epithelial cells. The immunized mouse spleen cells were fused with myeloma cell line NS-1. The McAb against human lens epithelial cell was screened by indirect immunofluorescence techniques. Its specificity was confirmed both by immunohistochemical analysis and by indirect immunofluorescence techniques. The large quantities of McAb were prepared by the hybridoma cells inoculated to the peritoneal cavity of BALB/c mouse. The McAb was purified by hydroxyapatite chromatography.
Results: One hybridoma cell line secreting anti-human lens epithelial cell McAb was successfully obtained, its subclass is IgM. Both immunohistochemical and immunofluorescence examinations demonstrated that the McAb was only positive to human lens epithelial cell membrane and negative to the other tissues of human eyeball. The mouse ascitic fluid was purified by hydroxyapatite chromatography and the purity was up to mass fraction 90%.
Conclusions: The McAb against human lens epithelial cell was successfully prepared, which can serve as a carrier of the target-directing agents designated to actively identify and selectively kill human lens epithelial cells. The agents may be more efficient in prevention and treatment of posterior capsular opacification (PCO).
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