2. Academic Validation
  3. Development of a High-Quality ELISA Method for Dinotefuran Based on a Novel and Newly-Designed Antigen

Development of a High-Quality ELISA Method for Dinotefuran Based on a Novel and Newly-Designed Antigen

  • Molecules. 2019 Jul 2;24(13):2426. doi: 10.3390/molecules24132426.
Fangfang Zhao 1 2 3 4,  Jingkun Liu 5,  Jinhui Luo 6 7 8 9
Affiliations

Affiliations

  • 1 Analysis & Testing Center, Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, Hainan, China. [email protected].
  • 2 Laboratory of Quality & Safety Risk Assessment for Tropical Products(Haikou), Ministry of Agriculture, Haikou 571101, Hainan, China. [email protected].
  • 3 Hainan Provincial Key Laboratory of Quality and Safety for Tropical Fruits and Vegetables, Haikou 571101, Hainan, China. [email protected].
  • 4 Quality Supervision and Inspection Center of Tropical Agro-Products, Haikou 571101, Hainan, China. [email protected].
  • 5 Institute of Environment and Plant Protection, Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, Hainan, China.
  • 6 Analysis & Testing Center, Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, Hainan, China.
  • 7 Laboratory of Quality & Safety Risk Assessment for Tropical Products(Haikou), Ministry of Agriculture, Haikou 571101, Hainan, China.
  • 8 Hainan Provincial Key Laboratory of Quality and Safety for Tropical Fruits and Vegetables, Haikou 571101, Hainan, China.
  • 9 Quality Supervision and Inspection Center of Tropical Agro-Products, Haikou 571101, Hainan, China.
PMID: 31269647 DOI: 10.3390/molecules24132426
Abstract

The structure of hapten determines the performance of the antibody and the corresponding detection method. A new type of antigen was designed and synthesized to expose the spatial and characteristic structure of dinotefuran molecule, and a type of high-quality antibody was obtained. The IC50 value of the monoclonal antibody was 5.30 ng/mL and its cross-reactivity (CRs) was less than 2% when reacting with other structurally related analytes. The effects of spatial configurations of hapten on the antibody were visually analyzed while using the appropriate software according to the quality of the Antibodies, which showed that the specificity of the antibody is closely related with the exposed structure of hapten. An ELISA assay with an IC50 of 5.66 ng/mL and a linear range of 1.95 to 16.29 ng/mL was developed. The results that were obtained from the ELISA and HPLC methods were equivalent. The results showed that spatial simulation is a crucial method that is used in the designing of hapten to obtain a sensitive and specific antibody. The application of this method will highlight the potential aim and improve the detection efficiency of ELISA.

Keywords

ELISA; Hapten design; dinotefuran; sensitivity and specificity; spatial configurations.

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