title: Multiplexed analysis of glycan variation on native proteins captured by antibody microarrays.
abstract: Carbohydrate post-translational modifications on proteins are important determinants of protein function in both normal and disease biology. We have developed a method to allow the efficient, multiplexed study of glycans on individual proteins from complex mixtures, using antibody microarray capture of multiple proteins followed by detection with lectins or glycan-binding antibodies. Chemical derivatization of the glycans on the spotted antibodies prevented lectin binding to those glycans. Multiple lectins could be used as detection probes, each targeting different glycan groups, to build up lectin binding profiles of captured proteins. By profiling both protein and glycan variation in multiple samples using parallel sandwich and glycan-detection assays, we found cancer-associated glycan alteration on the proteins MUC1 and CEA in the serum of pancreatic cancer patients. Antibody arrays for glycan detection are highly effective for profiling variation in specific glycans on multiple proteins and should be useful in diverse areas of glycobiology research.
authors.affiliation: Van Andel Research Institute, 333 Bostwick, Grand Rapids, Michigan 49503, USA.
authors.forename: Songming
authors.initials: S
authors.lastname: Chen
authors.forename: Tom
authors.initials: T
authors.lastname: LaRoche
authors.forename: Darren
authors.initials: D
authors.lastname: Hamelinck
authors.forename: Derek
authors.initials: D
authors.lastname: Bergsma
authors.forename: Dean
authors.initials: D
authors.lastname: Brenner
authors.forename: Diane
authors.initials: D
authors.lastname: Simeone
authors.forename: Randall E
authors.initials: RE
authors.lastname: Brand
authors.forename: Brian B
authors.initials: BB
authors.lastname: Haab
mesh_terms: D000906:Antibodies / Q000737:chemistry*; D000951:Antigens, Neoplasm / Q000737:chemistry*; D002272:Carcinoembryonic Antigen / Q000737:chemistry*; D006801:Humans; D037102:Lectins / Q000737:chemistry*; D046228:Microarray Analysis / Q000379:methods*; D018396:Mucin-1; D009077:Mucins / Q000737:chemistry*; D010190:Pancreatic Neoplasms / Q000097:blood / Q000378:metabolism*; D011134:Polysaccharides / Q000737:chemistry* / Q000276:immunology*; D011499:Protein Processing, Post-Translational; D014168:Transferrin / Q000737:chemistry*
chemical_list: D000906:Antibodies; D000951:Antigens, Neoplasm; D002272:Carcinoembryonic Antigen; D037102:Lectins; C060192:MUC1 protein, human; D018396:Mucin-1; D009077:Mucins; D011134:Polysaccharides; D014168:Transferrin