Investigating Colorectal Carcinoma Dependent Changes in Fucosylation of Carcinoembryogenic Antigen via Electron Transfer Dissociation in Tandem Mass Spectrometry

Abstract

In this research proposal I have written down a strategy to analyse the fucosylation pattern of the glycoprotein carcinoembryogenic antigen (CEA) originating from colorectal carcinoma (CRC) tissue and compare this to the fucosylation pattern of CEA originating from colorectal tissue of healthy individuals. The reason for investigating this is that many human fucosyltransferases are upregulated in CRC. Investigating the effect of this on CEA glycosylation could lead to new insights and medical applications regarding CRC. The primary method of analysis will be tandem mass spectrometry (MS) utilizing collision induced dissociation (CID) and electron transfer dissociation (ETD) as fragmentation methods. Several techniques will be used to isolate fucosylated structures, such as lectin affinity chromatography, facilitating analysis by MS. By analysing not only the amount of fucose present but also its different bonding conformations and the differences therein between different glycosylation sites on the protein – developing a specific and sensitive screening method for CRC might be possible. Gaining this detailed structural information is made possible by the use of ETD which has not been applied for these purposes before on glycans of this level of complexity. The diagnostic test that is the ultimate goals of this research would be in the form of a blood test which is less invasive than conventional CRC screening methods and would lead to more regular testing across the population, ultimately leading to significantly less CRC deaths.