Although there is ample evidence that the advanced glycation end-product (AGE) glucosepane contributes to age-related morbidities and diabetic complications, the impact of glucosepane modifications on proteins has not been extensively explored due to the lack of sufficient analytical tools. Here, we report the development of the first polyclonal anti-glucosepane antibodies using a synthetic immunogen that contains the core bicyclic ring structure of glucosepane. We investigate the recognition properties of these antibodies through ELISAs involving an array of synthetic AGE derivatives and determine them to be both high-affinity and selective in binding glucosepane. We then employ these antibodies to image glucosepane in aging mouse retinae via immunohistochemistry. Our studies demonstrate for the first time accumulation of glucosepane within the retinal pigment epithelium, Bruch’s membrane, and choroid: all regions of the eye impacted by age-related macular degeneration. Co-localization studies further suggest that glucosepane colocalizes with lipofuscin, which has previously been associated with lysosomal dysfunction and has been implicated in the development of age-related macular degeneration, among other diseases. We believe that the anti-glucosepane antibodies described in this study will prove highly useful for examining the role of glycation in human health and disease.

中文翻译：

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能够直接检测视网膜组织中葡萄糖烷的抗葡萄糖烷抗体的生成和表征

尽管有充分的证据表明晚期糖基化终产物（AGE）葡萄糖烷会导致与年龄相关的发病率和糖尿病并发症，但由于缺乏足够的分析工具，葡萄糖烷修饰对蛋白质的影响尚未得到广泛探索。在这里，我们报告了使用含有葡萄糖庚烷核心双环结构的合成免疫原开发出第一个多克隆抗葡萄糖庚烷抗体。我们通过涉及一系列合成 AGE 衍生物的 ELISA 研究了这些抗体的识别特性，并确定它们在结合葡聚糖方面具有高亲和力和选择性。然后，我们利用这些抗体通过免疫组织化学对衰老小鼠视网膜中的葡萄糖进行成像。我们的研究首次证明葡萄糖烷在视网膜色素上皮、布鲁赫膜和脉络膜内积累：所有受年龄相关性黄斑变性影响的眼睛区域。共定位研究进一步表明，葡萄糖烷与脂褐素共定位，脂褐素先前被认为与溶酶体功能障碍有关，并且与年龄相关性黄斑变性等疾病的发生有关。我们相信，本研究中描述的抗葡聚糖抗体对于检查糖化在人类健康和疾病中的作用将非常有用。