Development of the GlycoFibrotyper for detection of liver fibrosis

开发用于检测肝纤维化的 GlycoFibrotyper

• 批准号: 9909123
• 负责人: Anand S. Mehta
• 金额: $ 22.31万
• 依托单位: GLYCOPATH, INC
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-17 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9909123
• 关键词: Acute Disease; Antibodies; Binding; Binding Proteins; Biological Assay; Biological Markers; Capillary Electrophoresis; Chromatography; Chronic Disease; Cirrhosis; Classification; Clinical; Detection; Development; Diagnostic; Digestive System Disorders; Disease; Enzyme-Linked Immunosorbent Assay; Fibrosis; Fluorescent Dyes; Glycoproteins; Goals; Heart Diseases; Hepatitis C virus; IgG1; IgG2; IgG3; IgG4; Image; Immunoglobulin A; Immunoglobulin G; Immunoglobulin M; Immunoglobulins; Individual; Label; Laboratories; Lectin; Link; Liver; Liver Cirrhosis; Liver Fibrosis; Liver diseases; MALDI-TOF Mass Spectrometry; Malignant - descriptor; Malignant Neoplasms; Methods; Modification; Parents; Patients; Peptide N-glycohydrolase F; Phase; Plasma; Polysaccharides; Population; Precancerous Conditions; Premalignant; Primary carcinoma of the liver cells; Reagent; Reporting; Reproducibility; Resources; Rheumatoid Arthritis; Sampling; Serum; Slide; Small Business Technology Transfer Research; Specificity; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Testing; Time; Validation; Work; base; cohort; glycosylation; non-alcoholic fatty liver disease; nonalcoholic steatohepatitis; sugar

Increasingly, alterations in N-linked glycans have been reported for serum or plasma, or for the most abundant serum glycoprotein, immunoglobulin G, from large cohorts of samples representing rheumatoid arthritis, digestive diseases, cancer and liver fibrosis. In the case of fibrosis, two tests have identified alterations in N- linked glycosylation on both total IgG populations and on specific IgG molecules. Both of these tests can detect significant fibrosis with a high degree of accuracy and are also able to detect intermediate levels of fibrosis. However, both tests have drawbacks in that they require specialized sample handling resources, extensive processing and purification prior to analysis, and are expensive in regards to reagents and processing. Our group has recently developed a streamlined antibody capture slide array approach to directly profile N-glycans of captured serum glycoproteins like IgG, a method requiring a few microliters of sample and simplified processing workflows that require no purification or sugar modifications prior to analysis. This parent 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, such as the Bruker Tissuetyper MALDI-TOF, which is already available in clinical laboratories. We hypothesize that this method can be used to identify glycan biomarkers reflective of the changes that occur during the development of many diseases, including liver fibrosis/cirrhosis as performed here.

越来越多地报告了血清或血浆中N-连接聚糖的改变，或最丰富的 血清糖蛋白，免疫球蛋白G，来自代表类风湿性关节炎的大量样本， 消化系统疾病、癌症和肝纤维化。在纤维化的情况下，两种测试已经确定了N- 总IgG群体和特异性IgG分子上的连接糖基化。这两种测试都能检测出 以高准确度检测显著的纤维化，并且还能够检测中间水平的纤维化。 然而，这两种测试都有缺点，因为它们需要专门的样品处理资源、大量的 在分析之前进行处理和纯化，并且在试剂和处理方面昂贵。 我们的团队最近开发了一种简化的抗体捕获载玻片阵列方法， 捕获的血清糖蛋白（如IgG）的N-聚糖，这是一种需要几微升样品和简化的方法。 在分析前不需要纯化或糖修饰的加工流程。这个父方法是 称为GlycoTyper。在该方法中，N-连接的聚糖从捕获的抗体释放， 糖蛋白，并通过MALDI-TOF质谱法直接分析，例如Bruker Tissuetyper MALDI-TOF，它已经在临床实验室中可用。我们假设这种方法可以用来 鉴定反映许多疾病发展过程中发生的变化的聚糖生物标志物， 包括肝纤维化/肝硬化。