title: Nanoparticle decorated surfaces with potential use in glycosylation analysis.
abstract: A majority of all biologically active proteins are glycosylated and various diseases have proven to correlate with alterations in protein glycosylation. Sensitive identification of different glycoprotein glycoforms is therefore of great diagnostic value. Here we describe a method with potential for glycoprotein profiling, based on lectins as capture probes immobilized on particulate substrates in the nm-range. The nanoparticles present high concentrations of attachment sites for specific ligands and cause minimal steric hindrance to binding. In the present model study the mannose-binding lectin ConA has been coupled to polystyrene nanoparticles via a poly(ethyleneoxide) linker which protects the protein conformation and activity and prevents unspecific protein adsorption. The ConA-coated particles are accommodated at different spots on the analytical surface via oligonucleotide linkage. This attachment, which relies on the hybridization of complementary oligonucleotides, allows firm fixation of the particles at specific positions. The ConA attached to the particles has retained conformation and activity and binds selectively to a series of different glycoproteins. The results indicate the potential for using a multi-lectin nanoparticle array in glycoprotein mapping.
authors.affiliation: Department of Surface Biotechnology, Uppsala University, Box 577, 751 23 Uppsala, Sweden. karin.fromell@ybioteknik.uu.se
authors.forename: Karin
authors.initials: K
authors.lastname: Fromell
authors.forename: Margaretha
authors.initials: M
authors.lastname: Andersson
authors.forename: Karine
authors.initials: K
authors.lastname: Elihn
authors.forename: Karin D
authors.initials: KD
authors.lastname: Caldwell
mesh_terms: D002458:Cell Fractionation; D003208:Concanavalin A / Q000737:chemistry*; D006023:Glycoproteins / Q000737:chemistry; D006031:Glycosylation; D049329:Nanostructures / Q000737:chemistry*; D020442:Poloxamer / Q000737:chemistry; D040081:Protein Array Analysis / Q000295:instrumentation / Q000379:methods; D013499:Surface Properties
chemical_list: D006023:Glycoproteins; D020442:Poloxamer; D003208:Concanavalin A