title: High-affinity immobilization of proteins using biotin- and GST-based coupling strategies.
abstract: Surface plasmon resonance (SPR) is a highly sensitive method for the detection of molecular interactions. One interacting partner is immobilized on the sensor chip surface while the other is injected across the sensor surface. This chapter focuses on high-affinity immobilization of protein substrates for affinity and kinetic analyses using biotin/streptavidin interaction and GST/anti-GST-antibody interaction.
authors.affiliation: Department of Biochemistry and Biophysics, The University of North Carolina, Chapel Hill, NC, USA.
authors.forename: Stephanie Q
authors.initials: SQ
authors.lastname: Hutsell
authors.forename: Randall J
authors.initials: RJ
authors.lastname: Kimple
authors.forename: David P
authors.initials: DP
authors.lastname: Siderovski
authors.forename: Francis S
authors.initials: FS
authors.lastname: Willard
authors.forename: Adam J
authors.initials: AJ
authors.lastname: Kimple
mesh_terms: D000595:Amino Acid Sequence; D055910:Antibodies, Immobilized / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D001710:Biotin / Q000737:chemistry / Q000378:metabolism*; D019921:Biotinylation; D005982:Glutathione Transferase / Q000276:immunology* / Q000378:metabolism; D055767:Immobilized Proteins / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism*; D007700:Kinetics; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000276:immunology; D019809:Streptavidin / Q000737:chemistry / Q000378:metabolism; D020349:Surface Plasmon Resonance / Q000379:methods*
chemical_list: D055910:Antibodies, Immobilized; D055767:Immobilized Proteins; D010455:Peptides; D011993:Recombinant Fusion Proteins; D001710:Biotin; D019809:Streptavidin; D005982:Glutathione Transferase