Advanced glycation end-products (AGEs) are a heterogeneous group of compounds formed by non-enzymatic reaction between reducing-sugar and Arg/Lys in proteins and are involved in various diabetic complications. GA-pyridine is derived from glycolaldehyde and is one of the most cytotoxic AGEs. Here, we established a single-chain Fv (scFv) antibody against GA-pyridine, 73MuL9-scFv, and examined the details of its specificity and antigen recognition by using various techniques involving biophysics, chemical biology and structural biology. We also synthesized several compounds that differ slightly in regard to the position and number of GA-pyridine substituent groups, and revealed that GA-pyridine was specifically bound to 73MuL9-scFv. Thermodynamic analysis revealed that the association of GA-pyridine to 73MuL9-scFv was an exothermic and enthalpy driven reaction, and thus that the antigen recognition involved multiple specific interactions. Crystallographic analysis of the Fv fragment of 73MuL9-scFv revealed that several CH-π and hydrogen bond interactions took place between the Fv-fragment and GA-pyridine, which was consistent with the results of thermodynamic analysis. Further studies using 73MuL9-scFv as a tool to clarify the relevance of GA-pyridine to diabetic complications are warranted.

中文翻译：

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对 GA-吡啶特异的单链 Fv 抗体的分子识别，GA-吡啶是一种高级糖基化终产物 (AGE)，使用生物物理技术和合成抗原类似物进行了阐明

晚期糖基化终产物 (AGEs) 是由蛋白质中的还原糖和 Arg/Lys 之间的非酶促反应形成的一组异质化合物，与各种糖尿病并发症有关。GA-吡啶衍生自乙醇醛，是最具细胞毒性的 AGE 之一。在这里，我们建立了针对 GA-吡啶的单链 Fv (scFv) 抗体 73MuL9-scFv，并通过使用涉及生物物理学、化学生物学和结构生物学的各种技术检查了其特异性和抗原识别的细节。我们还合成了几种在 GA-吡啶取代基的位置和数量方面略有不同的化合物，并揭示了 GA-吡啶与 73MuL9-scFv 特异性结合。热力学分析表明，GA-吡啶与 73MuL9-scFv 的结合是放热和焓驱动的反应，因此抗原识别涉及多种特异性相互作用。73MuL9-scFv的Fv片段的晶体学分析表明，Fv片段和GA-吡啶之间发生了几个CH-π和氢键相互作用，这与热力学分析结果一致。需要进一步研究使用 73MuL9-scFv 作为工具来阐明 GA-吡啶与糖尿病并发症的相关性。这与热力学分析的结果一致。需要进一步研究使用 73MuL9-scFv 作为工具来阐明 GA-吡啶与糖尿病并发症的相关性。这与热力学分析的结果一致。需要进一步研究使用 73MuL9-scFv 作为工具来阐明 GA-吡啶与糖尿病并发症的相关性。