Enzyme Cascade Amplification-Based Immunoassay Using Alkaline Phosphatase-Linked Single-Chain Variable Fragment Fusion Tracer and MnO2 Nanosheets for Detection of Deoxynivalenol in Corn Samples
Guifang Xie, Fujing Mao, Yirui Huang, Li Wen, Zhichang Sun, Zhenyun He, Xing Liu
TL;DR
A new immunoassay method using enzyme amplification and nanosheets is developed to detect a harmful cereal toxin called deoxynivalenol in corn samples.
Contribution
A novel enzyme cascade amplification immunoassay using scFv-ALP and MnO2 nanosheets for sensitive DON detection is introduced.
Findings
The assay achieved a detection limit of 0.45 ng/mL and a linear range of 1.2–35.41 ng/mL.
The method showed good accuracy in recovery experiments and high selectivity for deoxynivalenol.
Results from corn samples correlated well with high-performance liquid chromatography data.
Abstract
Deoxynivalenol (DON) is a common mycotoxin that contaminates cereals. Therefore, the development of sensitive and efficient detection methods for DON is essential to guarantee food safety and human health. In this study, an enzyme cascade amplification-based immunoassay (ECAIA) using a dual-functional alkaline phosphatase-linked single-chain fragment variable fusion tracer (scFv-ALP) and MnO2 nanosheets was established for DON detection. The scFv-ALP effectively catalyzes the hydrolysis of ascorbyl-2-phosphate (AAP) to produce ascorbic acid (AA). This AA subsequently interacts with MnO2 nanosheets to initiate a redox reaction that results in the loss of oxidizing properties of MnO2. In the absence of ALP, MnO2 nanosheets can oxidize 3,3′,5,5′-tetramethylbenzidine (TMB) to produce the blue oxidized product of TMB, which exhibits a signal at a wavelength of 650 nm for quantitative…
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Taxonomy
TopicsAdvanced biosensing and bioanalysis techniques · Advanced Nanomaterials in Catalysis · Biosensors and Analytical Detection