title: Thiol-mediated anchoring of ligands to self-assembled monolayers for studies of biospecific interactions.
abstract: We report a method to immobilize thiol-containing ligands onto self-assembled monolayers (SAMs) of alkanethiolates presenting chloracetylated hexa(ethylene glycol) groups. The chloroacetyl groups react with thiols under mild basic conditions, enabling the stable immobilization of biologically active ligands in a well-defined orientation. These SAMs on gold are well suited for studies of biospecific interactions of immobilized ligands with proteins and cells. As a demonstration, we functionalized these SAMs with thiol-containing derivatives of biotin and benzene sulfonamide and observed the specific binding of neutravidin and carbonic anhydrase, respectively. We also used this method to generate mixed SAMs presenting the Arg-Gly-Asp (RGD) peptide sequence and demonstrated the integrin-mediated adhesion of fibroblast cells to these SAMs. This approach would allow the immobilization of proteins and other sensitive biomolecules and ligands for a wide variety of applications in biotechnology.
authors.affiliation: The Howard P. Isermann Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Troy, New York, 12180, USA.
authors.forename: Kunal V
authors.initials: KV
authors.lastname: Gujraty
authors.forename: Randolph
authors.initials: R
authors.lastname: Ashton
authors.forename: Sridhar R
authors.initials: SR
authors.lastname: Bethi
authors.forename: Sandesh
authors.initials: S
authors.lastname: Kate
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Faulkner
authors.forename: G Kane
authors.initials: GK
authors.lastname: Jennings
authors.forename: Ravi S
authors.initials: RS
authors.lastname: Kane
mesh_terms: D000473:Alkanes / Q000737:chemistry; D000818:Animals; D001360:Avidin / Q000737:chemistry; D001554:Benzene / Q000737:chemistry; D001709:Biotechnology / Q000379:methods; D001710:Biotin / Q000737:chemistry; D002256:Carbonic Anhydrases / Q000737:chemistry; D005347:Fibroblasts / Q000378:metabolism; D008024:Ligands; D051379:Mice; D041681:NIH 3T3 Cells; D009842:Oligopeptides / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*; D013438:Sulfhydryl Compounds / Q000737:chemistry*; D013449:Sulfonamides / Q000737:chemistry
chemical_list: D000473:Alkanes; D008024:Ligands; D009842:Oligopeptides; D011506:Proteins; D013438:Sulfhydryl Compounds; D013449:Sulfonamides; C093483:neutravidin; D001360:Avidin; D001710:Biotin; C047981:arginyl-glycyl-aspartic acid; D002256:Carbonic Anhydrases; D001554:Benzene