title: Highly selective and sensitive detection of β-agonists using a surface plasmon resonance sensor based on an alkanethiol monolayer functionalized on a Au surface.
abstract: Immunosensor surfaces for surface plasmon resonance (SPR) have been constructed using a functionalized succinimidyl propanethiol monolayer as a linker to immobilize β-agonist protein conjugates on a Au surface. Because β-agonist is a small molecule, an indirect competitive inhibition immunoassay was used for detection. The lowest detection limits for ractopamine and salbutamol were 10 ppt (10 pg mL(-1)) and 5 ppt (5 pg mL(-1)), respectively. The fabricated immunosensor surface can be used again for detection after regeneration in 0.1 M sodium hydroxide. It was found that the same sensor surface could be reused for performing over 100 rapid immunoreactions. Moreover, one immunosensing-regeneration cycle requires only 600 s. The fabricated immunosensor surfaces were characterized using SPR and scanning tunneling microscopy observation. In the kinetic study of the indirect competitive immunosensing inhibition, the affinity constant (K1) of salbutamol antibody was smaller than the K1 of ractopamine antibody. Compared to a previous study of clenbuterol detection, it was concluded that the high K1 was coupled with low sensitivity. In the selectivity study, both immunosensor surfaces provided >90% of confidence level for the specific detection of β-agonist compounds. The fabrication of highly selective and sensitive sensor surfaces for detecting β-agonist compounds was confirmed.
authors.affiliation: Division of Environmental Materials Science, Graduate School of Environmental Science and Section of Materials Science, Faculty of Environmental Earth Science, Hokkaido University, Sapporo 060-0810, Japan; Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Gadjah Mada, Sekip Utara Kotak Pos 21 BLS, Yogyakarta 55281, Indonesia.
authors.lastname: Suherman
authors.affiliation: New Business Development Office, USHIO INC., 6-1 Ohtemachi 2-chome, Chiyoda-ku, Tokyo 100-8150, Japan.
authors.forename: Kinichi
authors.initials: K
authors.lastname: Morita
authors.affiliation: Division of Environmental Materials Science, Graduate School of Environmental Science and Section of Materials Science, Faculty of Environmental Earth Science, Hokkaido University, Sapporo 060-0810, Japan. Electronic address: t_kawa@ees.hokudai.ac.jp.
authors.forename: Toshikazu
authors.initials: T
authors.lastname: Kawaguchi
mesh_terms: D000420:Albuterol / Q000276:immunology / Q000302:isolation & purification*; D000906:Antibodies / Q000737:chemistry / Q000276:immunology; D015374:Biosensing Techniques; D006046:Gold / Q000737:chemistry; D006801:Humans; D055767:Immobilized Proteins / Q000737:chemistry / Q000276:immunology; D007118:Immunoassay; D016252:Microscopy, Scanning Tunneling; D010627:Phenethylamines / Q000276:immunology / Q000302:isolation & purification*; D020349:Surface Plasmon Resonance
chemical_list: D000906:Antibodies; D055767:Immobilized Proteins; D010627:Phenethylamines; C060897:ractopamine; D006046:Gold; D000420:Albuterol
keywords: Indirect competitive inhibition immunoassay; Surface plasmon resonance; β-agonist compounds