Characterization of Anti-Advanced Glycation End Product Antibodies to Nonenzymatically Lysine-Derived and Arginine-Derived Glycated Products
Characterization of Anti-Advanced Glycation End Product Antibodies to Nonenzymatically Lysine-Derived and Arginine-Derived Glycated Products
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DOI：
10.1080/15321810903188136
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发表时间：
2009-01-01
影响因子：
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通讯作者：
Lim, Sabina
中科院分区：
文献类型：
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作者：
Choi, Yeong-Gon;Lim, Sabina
N-羧甲基赖氨酸（CML）和N-羧乙基赖氨酸（CEL）被称为晚期糖基化终产物（AGEs），已知它们是由牛血清白蛋白（BSA）和D-葡萄糖之间的非酶糖化产生的。本研究旨在鉴定抗AGE抗体（包括抗CML和抗CEL抗体）的免疫反应性。以AGE修饰的牛血清白蛋白（AGE-BSA）为免疫原，制备了抗AGE的多克隆IgG。在氨基酸糖基化分析中，抗AGEIgG能较强地检测BSA上形成的AGEs，至少部分能检测Lys和Arg上形成的AGEs，这是由于抗AGEIgG能与还原剂NaCNBH 3产生的Lys和Arg上的AGEs发生免疫反应，而免疫前血清不能。由于抗CML抗体也可以与AGE-BSA强烈反应，这表明CML是与BSA交联的主要非酶糖化产物。此外，CEL与BSA的可区分的聚合与BSA的CML聚合相关，尽管比CML弱并且不是通过Lys糖化分析产生的。这些结果表明，抗AGE抗体可有效检测LyS衍生和Arg衍生的AGE，并且CML和CEL可不同地聚合白蛋白作为AGE-BSA上存在的主要AGE。
N epsilon-(carboxymethyl)lysine (CML) and N epsilon-(carboxyethyl)lysine (CEL) termed advanced glycation end products (AGEs) are known to be produced by nonenzymatic glycation between bovine serum albumin (BSA) and D-glucose. This study is to characterize the immunoreactivity of anti-AGE antibodies including anti-CML and anti-CEL antibodies. Using AGE-modified BSA (AGE-BSA) as an immunogen, a polyclonal anti-AGE immunoglobulin G (IgG) was produced. The anti-AGE IgG could strongly detect AGEs formed on BSA, at least in part, AGEs produced on both residues Lys and Arg due to its immunoreaction with Lys-derived and Arg-derived AGEs produced by NaCNBH3, a reducing agent, in amino acid glycation analysis, but the pre-immune serum could not. As the anti-CML antibody could also strongly react with AGE-BSA, this suggests that CML is a major nonenzymatically glycated product cross-linked to BSA. Furthermore, CEL is associated with distinguishable polymerization of BSA from CML polymerization of BSA, though weaker than CML and was not produced by Lys glycation analysis. These results indicate that the anti-AGE antibody is effective for detecting both Lys-derived and Arg-derived AGEs, and CML and CEL distinctively polymerize albumin as major AGEs present on AGE-BSA.