title: Sensing lectin-glycan interactions using lectin super-microarrays and glycans labeled with dye-doped silica nanoparticles.
abstract: A new microarray platform, based on lectin super-microarrays and glycans labeled with dye-doped nanoparticles, has been developed to study glycan-lectin interactions. Glycan ligands were conjugated onto fluorescein-doped silica nanoparticles (FSNPs) using a general photocoupling chemistry to afford FSNP-labeled glycan probes. Lectins were printed on epoxy slides in duplicate sets to generate lectin super-microarrays where multiple assays could be carried out simultaneously in each lectin microarray. Thus, the lectin super-microarray was treated with FSNP-labeled glycans to screen for specific binding pairs. Furthermore, a series of ligand competition assays were carried out on a single lectin super-microarray to generate the dose-response curve for each glycan-lectin pair, from which the apparent affinity constants were obtained. Results showed 4-7 orders of magnitude increase in affinity over the free glycans with the corresponding lectins. Thus, the glycan epitope structures having weaker affinity than the parent glycans could be readily identified and analyzed from the lectin super-microarrays.
authors.affiliation: Department of Chemistry, Portland State University, P.O. Box 751, Portland, OR, 97207-0751, USA.
authors.forename: Xin
authors.initials: X
authors.lastname: Wang
authors.forename: Elena
authors.initials: E
authors.lastname: Matei
authors.forename: Lingquan
authors.initials: L
authors.lastname: Deng
authors.forename: Leonardus
authors.initials: L
authors.lastname: Koharudin
authors.forename: Angela M
authors.initials: AM
authors.lastname: Gronenborn
authors.forename: Olof
authors.initials: O
authors.lastname: Ramström
authors.forename: Mingdi
authors.initials: M
authors.lastname: Yan
mesh_terms: D015374:Biosensing Techniques; D005456:Fluorescent Dyes / Q000737:chemistry; D037102:Lectins / Q000302:isolation & purification* / Q000378:metabolism; D008024:Ligands; D053758:Nanoparticles / Q000737:chemistry*; D011134:Polysaccharides / Q000302:isolation & purification* / Q000378:metabolism; D040081:Protein Array Analysis; D012822:Silicon Dioxide / Q000737:chemistry; D013194:Staining and Labeling
chemical_list: D005456:Fluorescent Dyes; D037102:Lectins; D008024:Ligands; D011134:Polysaccharides; D012822:Silicon Dioxide