Development of a sensitive surface plasmon resonance immunosensor for detection of 2,4-dinitrotoluene with a novel oligo (ethylene glycol)-based sensor surface.
A surface plasmon resonance (SPR) immunosensor for detection of 2,4-dinitrotoluene (2,4-DNT), which is a signature compound of 2,4,6-trinitrotoluene-related explosives, was developed by using a novel oligo (ethylene glycol) (OEG)-based sensor surface. A rabbit polyclonal antibody against 2,4-DNT (anti-DNPh-KLH-400 antibody) was prepared, and the avidity for 2,4-DNT and recognition capability were investigated by indirect competitive ELISA. The sensor surface was fabricated by immobilizing a 2,4-DNT analog onto an OEG-based self-assembled monolayer formed on a gold surface via an OEG linker. The fabricated surface was characterized by Fourier-transform infrared-refractive absorption spectrometry (FTIR-RAS). The immunosensing of 2,4-DNT is based on the indirect competitive principle, in which the immunoreaction between the anti-DNPh-KLH-400 antibody and 2,4-DNT on the sensor surface was inhibited in the presence of free 2,4-DNT in solution. The limit of detection for the immunosensor, calculated as three times the standard deviation of a blank value, was 20 pg mL(-1), and the linear dynamic range was found to be between 1 and 100 ng mL(-1). Additionally, the fabricated OEG-based surface effectively prevented non-specific adsorption of proteins, and the specific response to anti-DNPh-KLH-400 antibody was maintained for more than 30 measurement cycles.
Nagatomo
Kazutaka
K
Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka-shi, Fukuoka 812-8581, Japan.
Kawaguchi
Toshikazu
T
Miura
Norio
N
Toko
Kiyoshi
K
Matsumoto
Kiyoshi
K
D000042:Absorption; D000818:Animals; D000906:Antibodies / Q000276:immunology; D001667:Binding, Competitive; D002417:Cattle; D004136:Dinitrobenzenes / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D053834:Explosive Agents / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D005260:Female; D006241:Haptens / Q000276:immunology; D007118:Immunoassay / Q000379:methods*; D008970:Molecular Weight; D011092:Polyethylene Glycols / Q000737:chemistry*; D011817:Rabbits; D017550:Spectroscopy, Fourier Transform Infrared; D020349:Surface Plasmon Resonance / Q000379:methods*; D013499:Surface Properties
D000906:Antibodies; D004136:Dinitrobenzenes; D053834:Explosive Agents; D006241:Haptens; D011092:Polyethylene Glycols; C016403:2,4-dinitrotoluene