title: Simple immobilization of antibody in organic/inorganic hybrid thin films for immunosensing.
abstract: Simple method to immobilize antibody in organic/inorganic hybrid thin films, prepared easily by using liquid phase deposition (LPD), was proposed to enhance the sensitivity, reproducibility and stability of immunosensing chips. The LPD method could embed a monoclonal antibody of C-reactive protein (Anti-CRP), which is mixed with poly(l-lysine) (PL) as an organic binder, on/in titanium oxide thin layer deposited onto a surface plasmon resonance (SPR) sensing chip (F(Anti-CRP-PL)), allowing simple immobilization of anti-CRP capable of CRP detection. As a control, anti-CRP was non-covalently immobilized after preparing the titanium oxide layer containing PL (SIF(Anti-CRP-PL)). Binding experiments of CRP on the prepared sensor chips were performed by SPR measurements. The CRP binding capacity of anti-CRP on F(Anti-CRP-PL) was 54% greater than that of anti-CRP on SIF (Anti-CRP-PL), suggesting that the immobilization method could retain good antigen-binding activity of anti-CRP. Reproducibility of the antigen-antibody interaction was estimated by SPR using 10mM HCl as the regeneration regent. CRP was repeatedly detected, and the detection limit was calculated to be 19.9ngmL(-1) (CV: 8.9%). By using this method, antibodies could be simply immobilized to various kinds of sensing chips for the development of immunosensing systems.
authors.affiliation: Graduate School of Engineering, Kobe University, 1-1, Kobe 657-8501, Japan.
authors.forename: Akiko
authors.initials: A
authors.lastname: Murata
authors.forename: Tooru
authors.initials: T
authors.lastname: Ooya
authors.forename: Toshifumi
authors.initials: T
authors.lastname: Takeuchi
mesh_terms: D000327:Adsorption; D000906:Antibodies / Q000737:chemistry* / Q000276:immunology; D000941:Antigens / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D015374:Biosensing Techniques / Q000295:instrumentation; D004867:Equipment Design; D019544:Equipment Failure Analysis; D007118:Immunoassay / Q000295:instrumentation*; D007287:Inorganic Chemicals / Q000737:chemistry*; D008567:Membranes, Artificial; D009930:Organic Chemicals / Q000737:chemistry*; D020349:Surface Plasmon Resonance / Q000295:instrumentation*
chemical_list: D000906:Antibodies; D000941:Antigens; D007287:Inorganic Chemicals; D008567:Membranes, Artificial; D009930:Organic Chemicals