title: N-Glycan profile analysis of transferrin using a microfluidic compact disc and MALDI-MS.
abstract: It has been known for a long time that diseases can be associated with changes to the glycosylation of specific proteins. This has been shown for cancer, immunological disorders, and neurodegenerative diseases. The possibility of using the glycosylation patterns of proteins as biomarkers for disease would be a great asset for clinical research or diagnosis. There is at present a lack of rapid, automated, and cost-efficient analytical techniques for the determination of the glycosylation of specific serum proteins. We have developed a method for determining the glycosylation pattern of proteins based on the affinity capture of a specific serum protein, the enzymatic release of the N-linked glycans, and the analysis of the glycan pattern using MALDI-MS. All sample preparation is performed in a disposable centrifugal microfluidic disc. The sample preparation is miniaturized, requiring only 1 μL of sample per determination, and automated with the possibility of processing 54 samples in parallel in 3.5 h. We have developed a method for the glycosylation pattern analysis of transferrin. The method has been tested on serum samples from chronic alcohol abusers and a control group. Also, a SIMCA model was created and evaluated to discriminate between the two groups.
authors.affiliation: Department of Environmental Sciences and Analytical Chemistry, Stockholm University, Svante Ahrrenius v 12, 10691, Stockholm, Sweden.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Quaranta
authors.affiliation: Department of Environmental Sciences and Analytical Chemistry, Stockholm University, Svante Ahrrenius v 12, 10691, Stockholm, Sweden.|Department of Genetics and Microbiology, Maria Curie-Sklodowska Univeristy, Lublin, Poland.
authors.forename: Anna
authors.initials: A
authors.lastname: Sroka-Bartnicka
authors.affiliation: Department of Environmental Sciences and Analytical Chemistry, Stockholm University, Svante Ahrrenius v 12, 10691, Stockholm, Sweden.
authors.forename: Erik
authors.initials: E
authors.lastname: Tengstrand
authors.affiliation: Department of Environmental Sciences and Analytical Chemistry, Stockholm University, Svante Ahrrenius v 12, 10691, Stockholm, Sweden. gunnar.thorsen@aces.su.se.
authors.forename: Gunnar
authors.initials: G
authors.lastname: Thorsén
mesh_terms: D001331:Automation; D001798:Blood Proteins / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D011134:Polysaccharides / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D014168:Transferrin / Q000737:chemistry*
chemical_list: D001798:Blood Proteins; D011134:Polysaccharides; D014168:Transferrin
keywords: Glycan pattern; Glycomics; MALDI-MS; Microfluidic; N-linked oligosaccharide; Transferrin