title: Reflectometric interference spectroscopy-based immunosensing using immobilized antibody via His-tagged recombinant protein A.
abstract: The proposed approach demonstrated in this study provides an immunosensing system based on reflectometric interference spectroscopy (RIfS) in combination with an antibody immobilization method using histidine-tagged recombinant protein A. Carboxymethyldextran (CMD) was immobilized on a 3-aminopropyltriethoxysilane-treated a silicon nitride-coated silicon wafer, followed by chelating histidine-tagged recombinant protein A with copper (II) ions. The CMD-layer was found to be advantageous in terms of not only immobilization of histidine-tagged recombinant protein A-mediated an antibody against myoglobin (anti-Myo) but also prevention of non-specific binding of myoglobin. Myoglobin was repeatedly detected, and the apparent detection limit was 0.1 μg mL(-1). The proposed RIfS-based protein sensing system, in conjunction with the easy preparation of silicon-based inexpensive immunosensing chips, is expected to be applicable for label-free optical detection for other proteins in various fields.
authors: Graduate School of Engineering, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe 657-8501, Japan.
authors: Hyung Woo
authors: HW
authors: Choi
authors: Graduate School of Engineering, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe 657-8501, Japan.
authors: Yasuhiko
authors: Y
authors: Sakata
authors: Graduate School of Engineering, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe 657-8501, Japan.
authors: Tooru
authors: T
authors: Ooya
authors: Graduate School of Engineering, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe 657-8501, Japan. Electronic address: takeuchi@gold.kobe-u.ac.jp.
authors: Toshifumi
authors: T
authors: Takeuchi
mesh_terms: D055910:Antibodies, Immobilized / Q000276:immunology*; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods*; D003300:Copper; D003911:Dextrans; D006639:Histidine / Q000235:genetics / Q000378:metabolism*; D009211:Myoglobin / Q000032:analysis / Q000276:immunology; D011437:Propylamines; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D012821:Silanes; D012825:Silicon; D017655:Silicon Compounds; D013057:Spectrum Analysis / Q000379:methods*
chemical_list: D055910:Antibodies, Immobilized; D003911:Dextrans; D009211:Myoglobin; D011437:Propylamines; D011994:Recombinant Proteins; D012821:Silanes; D017655:Silicon Compounds; D006639:Histidine; D003300:Copper; C014392:carboxymethyl dextran; C477625:amino-propyl-triethoxysilane; C032734:silicon nitride; D012825:Silicon
keywords: Antibody; Histidine-tagged recombinant protein A; Immunosensing; Immunosensor; Myoglobin; Reflectometric interference spectroscopy