Development of the GlycoHCCTyper for the early detection of HCC

长期以来，糖基化的改变与糖尿病的发展和进展有关。 许多类型的慢性和急性疾病。我们的小组（梅塔）是第一个执行 血清中的聚糖分析和对特定糖型进行蛋白质组学（糖蛋白质组学）。使用 通过这些方法，我们鉴定了一些糖基化改变的血清糖蛋白， 肝细胞癌，一种原发性肝癌。 然而，由于技术的限制，我们被迫要么检查每种蛋白质， 或者检查蛋白质池，而不能将特定聚糖连接到给定的 蛋白在某些情况下，我们对纯化的蛋白质进行结构聚糖分析， 提供了最好的“生物标志物”信息，但需要几天到几周的时间进行分析。或者，我们 可以放弃真正的结构信息，使用凝集素来确定是否只有一种特定的糖， 部分存在，并以更快速的方式进行分析。但是，这通常是 一次只针对一种蛋白质在所有这些情况下， 由于所使用的技术，糖蛋白减少了。 为了解决这一限制，GlycoPath最近开发了一种流线型抗体捕获载玻片 阵列方法直接分析捕获的血清糖蛋白上的N-连接聚糖。这个过程 只需要几微升的样品，并利用简单的方法，不需要蛋白质 在分析之前进行纯化或糖修饰。该方法被称为GlycoTyper。 在该方法中，N-连接的聚糖从抗体捕获的糖蛋白中释放，并被 通过MALDI-TOF质谱直接分析。我们假设这种方法可以 用于鉴定反映在发展过程中发生的变化的聚糖生物标志物， 肝细胞癌在第一阶段的STTR应用中，我们预计将开发一个 使用捕获的蛋白质的MALDI-MS的可再现和可翻译的工作流程， 检测肝细胞癌的存在。

Alterations in glycosylation have long been associated with the development and progression of many types of chronic and acute diseases. Our group (Mehta) was one of the first to perform glycan analysis in serum and perform proteomics on specific glycoforms (glycoproteomics). Using such methods, we identified a number of serum glycoproteins with altered glycosylation in hepatocellular carcinoma, a primary cancer of the liver. However, because of limitations in technology, we were forced to either examine each protein one at a time or examine pools of proteins without the ability to link a particular glycan to a given protein. In some situations, we performed structural glycan analysis on purified proteins which provided the best “biomarker” information, but took days to weeks for analysis. Alternatively, we could forgo true structural information and use lectins to determine if only one specific sugar moiety was present, and perform analysis in a more rapid manner. But again, this was generally done only on one protein at a time. In all of these situations, the biomarker potential of these glycoproteins was diminished because of the technology used. To address this limitation, GlycoPath has recently developed a streamlined antibody capture slide array approach to directly profile N-linked glycans on captured serum glycoproteins. This process requires only a few microliters of sample and utilizes simple methods that require no protein purification or sugar modifications prior to analysis. This method is referred to as the GlycoTyper. In this method, N-linked glycans are released from antibody captured glycoproteins and are directly analyzed by MALDI-TOF mass spectrometry. We hypothesize that this method can be used to identify glycan biomarkers reflective of the changes that occur during the development of hepatocellular carcinoma. In this Phase I STTR application we anticipate developing a reproducible and translatable workflow using MALDI-MS of captured proteins that can accurately detect the presence of hepatocellular carcinoma.