自制单克隆抗体检测糖尿病大鼠肾脏和心脏糖基化终产物

Self-Made Monoclonal Antibodies Detect Diabetic Rat Kidney and Heart Glycation End Products

目的为探讨糖基化终产物在糖尿病慢性并发症发生发展过程中的作用,采用本实验室所制备的7F、5F、8A三株单抗,检测高血压糖尿病大鼠肾脏和心脏糖基化终产物。方法Western blotting分析所制备单抗与对照物(人血清白蛋白、孵育人血清白蛋白和牛血清白蛋白)的交叉反应性及其针对的抗原表位。用所得单抗对糖尿病大鼠肾脏和心脏进行糖基化终产物的免疫组织化学分析。结果所得抗体与糖基化终产物结合能力和特异性较强,初步鉴别其抗原表位非羧甲基赖氨酸。在糖尿病大鼠肾脏和心脏探查到葡萄糖源性糖基化终产物,而正常对照组不明显。结论3株抗糖基化终产物单克隆抗体细胞株具较高抗原特异性,可定性显示组织中的糖基化终产物。

Aim To explore the role of advanced glycation end products（AGE） in pathogenesis and development of diabetic complications.Methods Indirect ELISA and Western blotting were used to analyse the immunological property of McAb and the epitope which would be combined with McAb.AGE in serum of human and aortas,renal,heart from diabetic rats were also detected by McAb.Results McAb reacted with AGE specially and was combined with non-carboxymethyl lysine（non-CML）.AGE in renal and heart of diabetic rats had been detected,but were not obviously detected in normal rats.Conclusion McAb（non-CML）reacted with AGE specially,and might be of value for AGE measurement.