角膜内皮抗原及其抗体的提纯

目的；提纯角膜内皮抗原（corneal enodothelial antigen,CEN-Ag）及抗角膜内皮抗原抗体（anti-corneal endothelial atibody,ACEN-Ab），为角膜移植排斥反尖的理论研究提供基础。方法：用猪角膜内皮做抗原，上皮做对照。经离子交换层及2B4．14．1单克隆抗体亲和层析制得纯化的CEN－Ag和角膜上皮抗原（corneal epithelial antigen,KCEP-Ag），用提纯的CEN－Ag和CEP－Ag分别免疫家兔，取血清经亲和层析得到ACEN－Ab和抗角膜上皮抗原抗体（anti-corneal epithelial antibod,ACEP-Ab）。结果：经聚丙烯酰胺凝胶电泳（SDS－PAGE）分析，抗原分子质量分别为43kD和53kD，相应的免疫球蛋白是IgG2和IgG3。结论：CEN－Ag和ACEN－Ab可以被提纯并从理论上证明，不同的角膜抗原和抗体穿透房水屏蔽的能力有所不同和角膜排斥反应的临床分型是正确的。

Purification of corneal endothelial antigen and anti-corneal endothelial antibody

Objective:To purify corneal endothelial antigen(CEN Ag) and anti corneal endothelial antibody(ACEN Ab).Methods:The corneal endothelium of the pig was used to made antigen,the epithelium was used as a control. CEN Ag and corneal epithelial antigen(CEP Ag) were purified by anion exchange chromatography and affinity chromatography with 2B4.14.1 monoclonal antibody. Rabbits were immunized with purified CEN Ag and CEP Ag. The serum,ACEN Ab,and the anti corneal epithelial antibody(ACEP Ab) were determined by affinity chromatography.Results:With sodium dodecyl sulphate polyacrylamide gel electrophoresis(SDS PAGE),the relative molecular weights of CEN Ag and CEP Ag were 43 kD and 53 kD,and their immunoglobulins were IgG 2 and IgG 3.Conclusion:The author purified CEN Ag and CEP Ag and assayed their molecular weight. This establishes a basis for biological therapy in rejection of keratoplasty.