酶联免疫吸附分析方法(ELISA)检测血浆中糖基化终极产物  

ELISA detects the plasma advanced glycation end products

作　　者：高岩[1] 周谨[1] 王兴宇[1] 惠汝太 

机构地区：[1]中国医学科学院中国协和医科大学心血管病研究所阜外心血管病医院中德分子医学研究室,北京市100037

出　　处：《中国分子心脏病学杂志》2004年第4期224-227,共4页Molecular Cardiology of China

基　　金：国家重点基础性研究项目 (G2 0 0 0 0 5690 1 )

摘　　要：目的　建立竞争性ELISA方法 ,测定血液中的高级糖化终产物的含量。方法　对兔及牛血浆白蛋白进行体外葡萄糖修饰 ,得到糖基化终极产物抗原 ;用兔此抗原免疫新西兰大白兔得到与其特异结合的抗体。结果　 1 经过TNBS方法及聚丙稀凝胶电泳鉴定修饰抗原 ,发现修饰前后蛋白携带自由氨基数至少 35 %已被葡萄糖修饰且分子量也发生变化。 2 免疫获得抗糖基化终极产物的多克隆抗体 ,并鉴定了此多抗只抗葡萄糖修饰的蛋白而不抗载体蛋白。3 用葡萄糖修饰的蛋白及其多克隆抗体制成ELISA试剂盒 ,批间、批内变异系数分别为 5 8%和 9 8% ,灵敏度为 0 5u/ml。结论　得到检测血浆糖基化终极产物的ELISA试剂盒。Objectives To establish the ELISA method to detect the plasma advanced glycation end products. Methods 1. AGEs were prepared in vitro by incubating rabbit （AGEs-RSA）and bovine albumin （AGEs- BSA）with glucose; Polyclonal Anti-AGEs antibodies were generated by injecting subcutaneously AGEs-RSA into New Zealand white rabbit. Results 1. At least 35 % of free amino groups of albumin was modified by glucose by using TNBS and Molecular weight before and after modifying have changed by using PAGE methods. 2. The polyclonal Anti-AGEs antibodies were specifically against AGEs-proteins not the carrier proteins. 3. The plasma AGEs levels was measured with ELISA by using the generated antibodies and modified albumin as standard and the interassay and intra-assay were 5.8% and 9.8%, respectively. Conclusion A stable ELISA method for detecting the plasma AGEs has been established.

关 键 词：糖基化终极产物(AGEs) ELISA 

分 类 号：R446.6[医药卫生—诊断学]