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.
Public Health Relevance Statement: Narrative: This application will develop a new platform for glycan analysis and determine its ability to detect liver fibrosis.
Project Terms: Acute Disease; Antibodies; base; Binding; Binding Proteins; Biological Assay; Biological Markers; Capillary Electrophoresis; Chromatography; Chronic Disease; Cirrhosis; Classification; Clinical; cohort; Detection; Development; Diagnostic; Digestive System Disorders; Disease; Enzyme-Linked Immunosorbent Assay; Fibrosis; Fluorescent Dyes; Glycoproteins; glycosylation; 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 diseases; Liver Fibrosis; MALDI-TOF Mass Spectrometry; Malignant - descriptor; Malignant Neoplasms; Methods; Modification; non-alcoholic fatty liver disease; nonalcoholic steatohepatitis; 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; sugar; Testing; Time; Validation; Work
