title: Development of a rapid method for the quantitative determination of deoxynivalenol using Quenchbody.
abstract: Quenchbody (Q-body) is a novel fluorescent biosensor based on the antigen-dependent removal of a quenching effect on a fluorophore attached to antibody domains. In order to develop a method using Q-body for the quantitative determination of deoxynivalenol (DON), a trichothecene mycotoxin produced by some Fusarium species, anti-DON Q-body was synthesized from the sequence information of a monoclonal antibody specific to DON. When the purified anti-DON Q-body was mixed with DON, a dose-dependent increase in the fluorescence intensity was observed and the detection range was between 0.0003 and 3 mg L(-1). The coefficients of variation were 7.9% at 0.003 mg L(-1), 5.0% at 0.03 mg L(-1) and 13.7% at 0.3 mg L(-1), respectively. The limit of detection was 0.006 mg L(-1) for DON in wheat. The Q-body showed an antigen-dependent fluorescence enhancement even in the presence of wheat extracts. To validate the analytical method using Q-body, a spike-and-recovery experiment was performed using four spiked wheat samples. The recoveries were in the range of 94.9-100.2%. The concentrations of DON in twenty-one naturally contaminated wheat samples were quantitated by the Q-body method, LC-MS/MS and an immunochromatographic assay kit. The LC-MS/MS analysis showed that the levels of DON contamination in the samples were between 0.001 and 2.68 mg kg(-1). The concentrations of DON quantitated by LC-MS/MS were more strongly correlated with those using the Q-body method (R(2) = 0.9760) than the immunochromatographic assay kit (R(2) = 0.8824). These data indicate that the Q-body system for the determination of DON in wheat samples was successfully developed and Q-body is expected to have a range of applications in the field of food safety.
authors: Division of Microbiology, National Institute of Health Sciences, 1-18-1, Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan.
authors: Tomoya
authors: T
authors: Yoshinari
authors: Biomedical Division, Ushio Inc., 1-12 Minamiwatarida-cho, Kawasaki-ku, Kawasaki 210-0855, Japan.
authors: Hiroyuki
authors: H
authors: Ohashi
authors: Biomedical Division, Ushio Inc., 1-12 Minamiwatarida-cho, Kawasaki-ku, Kawasaki 210-0855, Japan.
authors: Ryoji
authors: R
authors: Abe
authors: Biomedical Division, Ushio Inc., 1-12 Minamiwatarida-cho, Kawasaki-ku, Kawasaki 210-0855, Japan.
authors: Rena
authors: R
authors: Kaigome
authors: Research Center for Environmental Genomics, Kobe University, 1-1 Rokkodai, Nada, Kobe 657-8501, Japan.
authors: Hideo
authors: H
authors: Ohkawa
authors: Department of Food and Life Science, Azabu University, 1-17-71 Fuchinobe, Chuo-ku, Sagamihara, Kanagawa 252-5201, Japan. Electronic address: y-konishi@azabu-u.ac.jp.
authors: Yoshiko
authors: Y
authors: Sugita-Konishi
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D015374:Biosensing Techniques / Q000379:methods*; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry; D005506:Food Contamination / Q000032:analysis*; D005670:Fusarium / Q000737:chemistry*; D057230:Limit of Detection; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009183:Mycotoxins / Q000032:analysis*; D013050:Spectrometry, Fluorescence / Q000379:methods; D053719:Tandem Mass Spectrometry; D014255:Trichothecenes / Q000032:analysis*; D014908:Triticum / Q000737:chemistry / Q000382:microbiology*
chemical_list: D000911:Antibodies, Monoclonal; D005456:Fluorescent Dyes; D009183:Mycotoxins; D014255:Trichothecenes; C007262:deoxynivalenol
keywords: Deoxynivalenol; Quenchbody; Rapid analysis technique; Wheat