@article{27eb97e0d94e11dc860c000ea68e967b,
title = "Rapid and individual-specific glycoprofiling of the low abundance N-glycosylated protein tissue inhibitor of metalloproteinases-1",
abstract = "A gel-based method for a mass spectrometric site-specific glycoanalysis was developed using a recombinant glycoprotein expressed in two different cell lines. Hydrophilic interaction liquid chromatography at nanoscale level was used to enrich for glycopeptides prior to MS. The glycoprofiling was performed using matrix-assisted laser desorption/ionization MS and MS/MS. The method proved to be fast and sensitive and furthermore yielded a comprehensive site-specific glycan analysis, allowing a differentiation of the glycoprofiles of the two sources of recombinant protein, both comprising N-glycans of a highly heterogeneous nature. To test the potential of the method, tissue inhibitor of metalloproteinases-1 (TIMP-1), a secreted low abundance N-glycosylated protein and a cancer marker, was purified in an individual-specific manner from plasma of five healthy individuals using IgG depletion and immunoaffinity chromatography. The corresponding TIMP-1 glycoprofiles were determined to be highly similar, comprising mainly bi- and triantennary complex oligosaccharides. Additionally it was shown that platelet-derived TIMP-1 displayed a similar glycoprofile. This is the first study to investigate the glycosylation of naturally occurring human TIMP-1, and the high similarity of the glycoprofiles showed that individual-specific glycosylation variations of TIMP-1 are minimal. In addition, the results showed that TIMP-1 derived from platelets and plasma is similarly glycosylated. This comprehensive and rapid glycoprofiling of a low abundance glycoprotein performed in an individual-specific manner allows for future studies of glycosylated biomarkers for person-specific detection of altered glycosylation and may thus allow early detection and monitoring of diseases.",
keywords = "Amino Acid Sequence, Animals, Blood Platelets, CHO Cells, Cell Line, Chromatography, Affinity, Cricetinae, Cricetulus, Glycosylation, Humans, Peptide Fragments, Plasma, Protein Array Analysis, Proteomics, Recombinant Proteins, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Tandem Mass Spectrometry, Tissue Inhibitor of Metalloproteinase-1",
author = "Morten Thaysen-Andersen and Th{\o}gersen, \{Ida B\} and Nielsen, \{Hans J{\o}rgen\} and Ulrik Lademann and Nils Br{\"u}nner and Enghild, \{Jan J\} and Peter H{\o}jrup",
year = "2007",
month = apr,
day = "1",
doi = "10.1074/mcp.M600407-MCP200",
language = "English",
volume = "6",
pages = "638--647",
journal = "Molecular and Cellular Proteomics",
issn = "1535-9476",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "4",
}