Nakayama H, Kato M, Makita Z, Aoki S, Kuroda Y, Misawa K, Yanagisawa K, Nakagawa S, Ikeda K
2nd Department of Internal Medicine, Hokkaido University School of Medicine, Sapporo, Japan.
J Immunol Methods. 1988 Aug 9;112(1):57-61. doi: 10.1016/0022-1759(88)90033-6.
A competitive radioimmunoassay for an advanced glycosylation endproduct, 2-(2-furoyl)-4(5)-(2-furanyl)-1H-imidazole (FFI) was developed. The immunogen was prepared by coupling of 4-furanyl-2-furoyl-1H-imidazole-1-hexanoic acid to keyhole limpet hemocyanin. The antiserum obtained by immunizing guinea pigs with the immunogen exhibited high affinity binding to FFI, but no cross-reactivity was observed for structurally related compounds containing an imidazole ring or furan ring(s). By using the radioimmunoassay, the levels of FFI in bovine serum albumin incubated with glucose for varying lengths of time were measured. A time-dependent increase was obtained in the amount of acid-liberated FFI and fluorescence. The radioimmunoassay described here had satisfactory reproducibility as judged by the intra-assay precision of 3.4-6.4% and the interassay precision of 7.3-8.9%. The method allows to quantitate FFI on the modified proteins that have been implicated in the complications of diabetes and in normal aging as well.
开发了一种针对晚期糖基化终产物2-(2-呋喃甲酰基)-4(5)-(2-呋喃基)-1H-咪唑(FFI)的竞争性放射免疫测定法。免疫原是通过将4-呋喃基-2-呋喃甲酰基-1H-咪唑-1-己酸偶联到匙孔血蓝蛋白上制备的。用该免疫原免疫豚鼠获得的抗血清对FFI表现出高亲和力结合，但对含有咪唑环或呋喃环的结构相关化合物未观察到交叉反应。通过使用放射免疫测定法，测量了与葡萄糖孵育不同时间的牛血清白蛋白中FFI的水平。酸释放的FFI量和荧光呈时间依赖性增加。根据3.4-6.4%的批内精密度和7.3-8.9%的批间精密度判断，这里描述的放射免疫测定法具有令人满意的重现性。该方法能够对与糖尿病并发症以及正常衰老相关的修饰蛋白上的FFI进行定量。
