title: A versatile method for functionalizing surfaces with bioactive glycans.
abstract: Microarrays and biosensors owe their functionality to our ability to display surface-bound biomolecules with retained biological function. Versatile, stable, and facile methods for the immobilization of bioactive compounds on surfaces have expanded the application of high-throughput "omics"-scale screening of molecular interactions by nonexpert laboratories. Herein, we demonstrate the potential of simplified chemistries to fabricate a glycan microarray, utilizing divinyl sulfone (DVS)-modified surfaces for the covalent immobilization of natural and chemically derived carbohydrates, as well as glycoproteins. The bioactivity of the captured glycans was quantitatively examined by surface plasmon resonance imaging (SPRi). Composition and spectroscopic evidence of carbohydrate species on the DVS-modified surface were obtained by X-ray photoelectron spectroscopy (XPS) and time-of-flight secondary ion mass spectrometry (ToF-SIMS), respectively. The site-selective immobilization of glycans based on relative nucleophilicity (reducing sugar vs amine- and sulfhydryl-derived saccharides) and anomeric configuration was also examined. Our results demonstrate straightforward and reproducible conjugation of a variety of functional biomolecules onto a vinyl sulfone-modified biosensor surface. The simplicity of this method will have a significant impact on glycomics research, as it expands the ability of nonsynthetic laboratories to rapidly construct functional glycan microarrays and quantitative biosensors.
authors.affiliation: Department of Bioengineering, University of Washington, Seattle, Washington 98195, United States.
authors.forename: Fang
authors.initials: F
authors.lastname: Cheng
authors.forename: Jing
authors.initials: J
authors.lastname: Shang
authors.forename: Daniel M
authors.initials: DM
authors.lastname: Ratner
mesh_terms: D000586:Amination; D006878:Hydroxides / Q000737:chemistry; D011134:Polysaccharides / Q000737:chemistry* / Q000378:metabolism*; D013379:Substrate Specificity; D013450:Sulfones / Q000737:chemistry; D013499:Surface Properties
chemical_list: D006878:Hydroxides; D011134:Polysaccharides; D013450:Sulfones; C009873:divinyl sulfone; C031356:hydroxide ion