title: Photogenerated lectin sensors produced by thiol-ene/yne photo-click chemistry in aqueous solution.
abstract: The photoinitiated radical reactions between thiols and alkenes/alkynes (thiol-ene and thiol-yne chemistry) have been applied to a functionalization methodology to produce carbohydrate-presenting surfaces for analyses of biomolecular interactions. Polymer-coated quartz surfaces were functionalized with alkenes or alkynes in a straightforward photochemical procedure utilizing perfluorophenylazide (PFPA) chemistry. The alkene/alkyne surfaces were subsequently allowed to react with carbohydrate thiols in water under UV-irradiation. The reaction can be carried out in a drop of water directly on the surface without photoinitiator, and any disulfide side products were easily washed away after the functionalization process. The resulting carbohydrate-presenting surfaces were evaluated in real-time studies of protein-carbohydrate interactions using a quartz crystal microbalance (QCM) flow-through system with recurring injections of selected lectins, with intermediate regeneration steps using low pH buffer. The resulting methodology proved fast, efficient and scalable to high-throughput analysis formats, and the produced surfaces showed significant protein binding with expected selectivities of the lectins used in the study.
authors.affiliation: KTH Royal Institute of Technology, Department of Chemistry, Stockholm, Sweden.
authors.forename: Oscar
authors.initials: O
authors.lastname: Norberg
authors.forename: Irene H
authors.initials: IH
authors.lastname: Lee
authors.forename: Teodor
authors.initials: T
authors.lastname: Aastrup
authors.forename: Mingdi
authors.initials: M
authors.lastname: Yan
authors.forename: Olof
authors.initials: O
authors.lastname: Ramström
mesh_terms: D000475:Alkenes / Q000737:chemistry; D000480:Alkynes / Q000737:chemistry; D001386:Azides / Q000737:chemistry*; D015374:Biosensing Techniques / Q000379:methods*; D002241:Carbohydrates / Q000737:chemistry; D057930:Click Chemistry / Q000379:methods*; D006845:Hydrocarbons, Fluorinated / Q000737:chemistry*; D037102:Lectins / Q000737:chemistry / Q000302:isolation & purification*; D010777:Photochemistry / Q000379:methods; D011108:Polymers / Q000737:chemistry; D011485:Protein Binding; D012996:Solutions; D013438:Sulfhydryl Compounds / Q000737:chemistry; D013499:Surface Properties
chemical_list: D000475:Alkenes; D000480:Alkynes; D001386:Azides; D002241:Carbohydrates; D006845:Hydrocarbons, Fluorinated; D037102:Lectins; D011108:Polymers; D012996:Solutions; D013438:Sulfhydryl Compounds; C556136:perfluorophenyl azide