Tn antigen (CD175), recognized as the precursor monosaccharide (α-GalNAc) of mucin O-glycan, is a well-known tumor-associated carbohydrate antigen (TACA). It has emerged as a potential biomarker for cancer diagnosis and prognosis. However, the role it plays in cancer biology remains elusive due to the absence of a sensitive and selective detection method. In this study, we synthesized two new probes based on a unique uridine-5′-diphospho-α-d-galactose (UDP-Gal) derivative, each functionalized with either a fluorescence or a cleavable biotin tag, to develop an innovative one-step enzymatic labeling strategy, enabling the visualization, enrichment, and site-specific mapping of the Tn antigen with unparalleled sensitivity and specificity. Our versatile strategy has been successfully applied to detect and image Tn antigen across various samples, including the complex cell lysates, live cells, serum, and tissue samples. Compared to the traditional lectin method, this one-step enzymatic method is simpler and more efficient (>10/100-fold in sensitivity). Furthermore, it allowed us to map 454 Tn-glycoproteins and 624 Tn-glycosylation sites from HEK293F^Tn+ and Jurkat cells. Therefore, our strategy provides an exceptionally promising tool for revealing the biological functions of the Tn antigen and advancing cancer diagnostics.

中文翻译：

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一种用于肿瘤相关 Tn 抗原高效成像和绘图的多功能一步酶策略


Tn 抗原 (CD175) 被认为是粘蛋白 O-聚糖的前体单糖 (α-GalNAc)，是一种众所周知的肿瘤相关碳水化合物抗原 (TACA)。它已成为癌症诊断和预后的潜在生物标志物。然而，由于缺乏灵敏和选择性的检测方法，它在癌症生物学中发挥的作用仍然难以捉摸。在这项研究中，我们合成了两种基于独特的尿苷-5'-二磷酸-α-d-半乳糖 (UDP-Gal) 衍生物的新探针，每种探针均带有荧光或可裂解的生物素标签，以开发一种创新的单-步骤酶标记策略，能够以无与伦比的灵敏度和特异性对 Tn 抗原进行可视化、富集和位点特异性定位。我们的多功能策略已成功应用于各种样品中的 Tn 抗原检测和成像，包括复杂的细胞裂解物、活细胞、血清和组织样品。与传统的凝集素方法相比，这种一步酶法更简单、更高效（灵敏度 >10/100 倍）。此外，它使我们能够从 HEK293F ^Tn+ 和 Jurkat 细胞中绘制出 454 个 Tn 糖蛋白和 624 个 Tn 糖基化位点。因此，我们的策略为揭示 Tn 抗原的生物学功能和推进癌症诊断提供了一种非常有前途的工具。