醣蛋白微陣列之發展與應用

Direct Screening of Glycan Patterns from Human Sera: A Selective Glycoprotein Microarray Strategy

杜秀中(Hsiu-Chung Tu) ； 林伯樵(Po-Chiao Lin)

《化學》 78卷1期 (2020/03) Pp. 83-91

https://doi.org/10.6623/chem.202003_78(1).007

摘要

蛋白質醣基化是複雜且重要的後轉譯修飾之一。其中，聚醣結構中的微小改變會影響癌細胞之變異及增生。因此，發展快速且直接的醣蛋白監測方法並能針對癌症中醣共軛體的過度分泌進行快速檢測，為現今社會重要挑戰之一。在此篇文章中，將介紹目前醣蛋白微陣列晶片的開發及應用。並在最後介紹本實驗室已發表無痕式探針亞硼酸－甲苯磺醯基在醣蛋白晶片之一系列開發應用。此策略能成功達到複雜環境中專一性醣基的辨識，並選擇性的針對醣基進行高專一性辨識。而近期搭配環張力疊氮炔環加成反應之晶片固化策略，能提升微陣列晶片於人體血清中醣蛋白之感測。此策略的建立促使我們對於人體血清中醣體模組的監測能有效掌握，並且在不經由繁瑣的樣品製備的條件下，就能比對健康人體以及臨床患者血清中的醣蛋白，並利用此無痕標記來作為區分癌症臨床診斷之重要平台。

關鍵字

微陣列晶片 ； 醣蛋白 ； 無痕標記 ； 亞硼酸－甲苯磺醯基

並列摘要

Protein glycosylation is one of the most complicated but significant post-translational modifications. Minor alterations in glycan structure can considerably affect the biology of a cell. Therefore, direct monitoring of glycan patterns of glycoproteins is closely related to cancer progression as well as metastasis. In this study, a boronic acid (BA)-tosyl-directed strategy to selectively immobilize glycoproteins on glass slides was successfully developed even in the presence of high-abundant nonglycosylated proteins. To enhance the immobilization efficiency and reduce the undesired nonspecific absorption, the strain-promoted alkyne azide cycloaddition (SPAAC) conjugation chemistry and surface blocking conditions were carefully optimized for the collection of reliable data. The optimized glycoprotein microarray platform describes specific lectin-recognition patterns of glycoproteins of interest in E. coil lysate and fetal bovine serum (FBS), which encourages us for direct monitoring of glycan patterns from human sera without tedious sample preparation. Three serum groups comprised of healthy controls and lung cancer and pancreatic cancer patients were analyzed by this new technique. Remarkably, the distinguishable glycan patterns of the three groups make them a powerful platform for cancer screening and prediagnosis.

並列關鍵字

Glycoprotein microarray ； SPAAC (strain-promoted alkyne azide cycloaddition) ； sensor ； lectin