摘要
目的探讨晚期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,对我们制备的5株抗糖基化终产物单克隆抗体进行特异性及抗原表位鉴定。方法采用间接酶联免疫吸附法和蛋白印迹术分析所制备单克隆抗体与对照物(人血清白蛋白、孵育人血清白蛋白、牛血清白蛋白)的交叉反应性及其针对的抗原表位。用所得单克隆抗体对正常对照、糖尿病及透析病人血清和糖尿病大鼠主动脉进行糖基化终产物的蛋白印迹分析。结果所得抗体与糖基化终产物特异性结合,而与对照物结合较低,提示它们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。在正常对照、糖尿病及透析病人血清中检测到葡萄糖源性糖基化终产物。用所得单克隆抗体行蛋白印迹分析,发现糖尿病高血压大鼠主动脉含糖基化终产物特异显色,而正常对照组不明显。结论5株抗糖基化终产物单克隆抗体细胞株具较高抗原特异性,可定性发现血清和组织中的糖基化终产物。
Aim To explore the role of advanced glycation end products （AGEs）in pathogenesis and development of diabetic complications, and analyse the immunological property of monodonal antibodies （McAb）against AGEs. Methods Indirect ELISA and western blotting were used to analyse the immunological property of McAb and the epitope which would combine with McAb. AGEs in serum of human and aortas from diabetic rats were also detected by McAb. Results McAb reacted with AGEs specially and combined with non-carboxymethyl lysine （ non-CML）. AGEs in serum of human and aortas from diabet- ic rats had been detected, however, there had been no AGEs detected in aortas from normal rats. Conclusion McAb（non- CML}reacted with AGEs specially,and might be of value for AGEs measurement.
出处
《中国动脉硬化杂志》
CAS
CSCD
2006年第5期409-412,共4页
Chinese Journal of Arteriosclerosis
