The aim of the project is to develop an immunoaffinity based assay for determining N-linked glycosylation profiles of specific serum proteins. The assay will use immobilized antibodies or similar affinity recognition molecules for capture of the proteins to be studied. Specific enzymes are used to release the glycans attached to the proteins of interest and the relative abundance of different glycans will be determined using Matrix Assisted Laser Desorption/Ionization (MALDI) Mass Spectrometry. Proteins connected to cancer, neurodegenerative diseases, or chronic pain will be used as target antigens and the long term goal of the project is to use the assay to generate mechanistic knowledge concerning the diseases or to increase the reliability of clinical assays through the ability of performing both quantitative analysis and glycoprofiling of the target proteins.
We have already developed automated and miniaturized microfluidic methods for N-linked glycan profiling of therapeutic antibodies [1]. The methods developed have been applied to the analysis of glycan patterns on therapeutic antibodies selectively captured from cell media supernatant [2] and serum samples [3]. The project has so far been performed in collaboration with Gyros AB.
