Development of the GlycoHCCTyper for the early detection of HCC

开发用于早期检测 HCC 的 GlycoHCCTyper

• 批准号: 10382624
• 负责人: Stephen Castellino
• 金额: $ 42.84万
• 依托单位: GLYCOPATH, INC
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-01 至 2024-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10382624
• 关键词: AFP gene; Acute Disease; Address; Algorithms; Antibodies; Biological Assay; Biological Markers; Blinded; Blood specimen; CA-125 Antigen; Ceruloplasmin; Chronic Disease; Cirrhosis; Clinical; Detection; Development; Diagnosis; Early Diagnosis; Ensure; Evaluation; FDA approved; Glycoproteins; Goals; Hand; Haptoglobins; Hemopexin; Image; Immunoglobulin G; Laboratories; Lectin; Link; Low-Molecular-Weight Kininogen; MALDI-TOF Mass Spectrometry; Malignant neoplasm of liver; Mass Spectrum Analysis; Medical; Methods; Modification; Orosomucoid; Patients; Phase; Polysaccharides; Primary Malignant Neoplasm of Liver; Primary carcinoma of the liver cells; Process; Proteins; Proteomics; Reproducibility; Sampling; Serum; Slide; Small Business Technology Transfer Research; South Carolina; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; System; Technology; Time; Tissue Sample; Transferrin; Universities; Validation; Vitamin D-Binding Protein; Work; alpha 2-Glucoproteins; alpha-Fetoproteins; apolipoprotein D; biomarker signature; cancer biomarkers; clinical development; cohort; design; glycoproteomics; glycosylation; histidine-rich glycoprotein; instrument; mass spectrometer; non-alcoholic fatty liver disease; nonalcoholic steatohepatitis; novel; prospective; protein purification; sample collection; sugar

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.

糖基化的改变长期以来一直与疾病的发生和进展有关 我们小组（Mehta）是最先进行治疗的小组之一。 血清中的聚糖分析并对特定糖型进行蛋白质组学（糖蛋白质组学）。 通过这种方法，我们鉴定了许多具有糖基化的血清糖蛋白 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 一次一个或检查蛋白质库，但无法将特定聚糖连接到给定的 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。 可以放弃真实的结构信息并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，但同样，这通常是。 在所有这些情况下，一次仅对一种蛋白质进行，这些生物标志物的潜力。 由于所使用的技术，糖蛋白减少了。 为了解决这一限制，GlycoPath 最近开发了一种简化的抗体捕获载玻片 阵列方法直接分析捕获的血清糖蛋白上的 N 连接聚糖。 仅需要几微升样品，并采用不需要蛋白质的简单方法 分析前的纯化或糖修饰此方法称为 GlycoTyper。 在此方法中，N-连接聚糖从抗体捕获的糖蛋白中释放出来，并被 我们发现该方法可以直接通过 MALDI-TOF 质谱进行分析。 用于识别反映发育过程中发生的变化的聚糖生物标志物 在此 I 期 STTR 应用中，我们预计开发一种肝细胞癌。 使用 MALDI-MS 捕获蛋白质的可重复和可翻译的工作流程，可以准确地 检测肝细胞癌的存在。