Establishment of a liquid chromatography-mass spectrometry method for the analysis of glycan biomarkers of pancreatic cancer

Abstract

Glycoproteins are proteins that have oligosaccharides covalently attached to the peptide backbone. These carbohydrates are also known as glycans. Protein glycans play a significant role in various cellular processes, including cell-cell recognition, signaling, and adhesion on cell surfaces. However, glycans undergo structural changes in many diseases, such as cancer. Therefore, there is a need to establish new analytical methods that aid in identifying and quantifying glycans to detect the presence of diseases. In the present study, a reference method for the separation and identification of labelled glycans using capillary liquid chromatography (capLC) coupled with ultraviolet (UV) and mass spectrometry (MS) detection will be developed. First, standard glycans selected as model will be derivatized with aniline (AN) and procainamide (ProA) labels. To assess their degree of derivatization, they will be analysed by matrix assisted laser desorption ionization mass spectrometry (MALDI-MS). Finally, these labelled glycans will serve for the development of a reference analytical method by capLC-UV and capLC-MS. This reference method will be used in the future to test the status of the chromatographic system in glycoprotein biomarker research. For instance, for the analysis of the N-glycans of human alpha-1-acid glycoprotein (hAGP) under study as a potential biomarker of pancreatic cancer.