title: Anti-idiotypic nanobody-alkaline phosphatase fusion proteins: Development of a one-step competitive enzyme immunoassay for fumonisin B1 detection in cereal.
abstract: A rapid and sensitive one-step competitive enzyme immunoassay for the detection of FB1 was developed. The anti-idiotypic nanobody-alkaline phosphatase (Ab2β-Nb-AP) was validated by the AP enzyme activity and the properties of bounding to anti-FB1-mAb (3F11) through colorimetric and chemiluminescence analyses. The 50% inhibitory concentration and the detection limit (LOD) of colorimetric enzyme-linked immunosorbent assay (ELISA) for FB1 were 2.69 and 0.35 ng mL(-1), respectively, with a linear range of 0.93-7.73 ng mL(-1). The LOD of the chemiluminescence ELISA (CLIA) was 0.12 ng mL(-1), and the IC50 was 0.89 ± 0.09 ng mL(-1) with a linear range of 0.29-2.68 ng mL(-1). Compared with LC-MS/MS, the results of this assay indicated the reliability of the Ab2β-Nb-AP fusion protein based one-step competitive immunoassay for monitoring FB1 contamination in cereals. The Ab2β-Nb-AP fusion proteins have the potential to replace chemically-coupled probes in competitive enzyme immunoassay systems.
authors.affiliation: State Key Laboratory of Food Science and Technology, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047, People's Republic of China; Jiangxi-OAI Joint Research Institute, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047, People's Republic of China.
authors.forename: Mei
authors.initials: M
authors.lastname: Shu
authors.affiliation: State Key Laboratory of Food Science and Technology, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047, People's Republic of China; Jiangxi-OAI Joint Research Institute, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047, People's Republic of China. Electronic address: xuyang@ncu.edu.cn.
authors.forename: Yang
authors.initials: Y
authors.lastname: Xu
authors.affiliation: State Key Laboratory of Food Science and Technology, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047, People's Republic of China; College of Food Science and Technology, Hainan University, No. 58 Renmin Avenue, Haikou 570228, China.
authors.forename: Xing
authors.initials: X
authors.lastname: Liu
authors.affiliation: Jiangxi-OAI Joint Research Institute, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047, People's Republic of China.
authors.forename: Yanping
authors.initials: Y
authors.lastname: Li
authors.affiliation: Jiangxi-OAI Joint Research Institute, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047, People's Republic of China.
authors.forename: Qinghua
authors.initials: Q
authors.lastname: He
authors.affiliation: State Key Laboratory of Food Science and Technology, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047, People's Republic of China.
authors.forename: Zhui
authors.initials: Z
authors.lastname: Tu
authors.affiliation: Jiangxi-OAI Joint Research Institute, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047, People's Republic of China.
authors.forename: Jinheng
authors.initials: J
authors.lastname: Fu
authors.affiliation: Department of Entomology and UCD Comprehensive Cancer Center, University of California, Davis, CA 95616, USA.
authors.forename: Shirley J
authors.initials: SJ
authors.lastname: Gee
authors.affiliation: Department of Entomology and UCD Comprehensive Cancer Center, University of California, Davis, CA 95616, USA.
authors.forename: Bruce D
authors.initials: BD
authors.lastname: Hammock
mesh_terms: D000469:Alkaline Phosphatase / Q000276:immunology*; D037341:Fumonisins / Q000276:immunology*; D007124:Immunoenzyme Techniques / Q000379:methods*; D061905:Single-Domain Antibodies / Q000276:immunology*
chemical_list: D037341:Fumonisins; D061905:Single-Domain Antibodies; C056933:fumonisin B1; D000469:Alkaline Phosphatase
keywords: Anti-idiotypic antibody; Nanobody; One-step immunoassay