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Citation: LI Xiao-Xiao, DING Shu-Mao, YANG Xu, YUAN Jun-Lin. Synthesis of Thifluzamide Artificial Antigens and Establishment of Enzyme Linked Immunosorbent Assay Detection Method[J]. Chinese Journal of Analytical Chemistry, ;2018, 46(1): 27-32. doi: 10.11895/j.issn.0253-3820.170206 shu

Synthesis of Thifluzamide Artificial Antigens and Establishment of Enzyme Linked Immunosorbent Assay Detection Method

Hubei Key Laboratory of Genetic Regulation and Integrative Biology, College of Life Science, Central China Normal University, Wuhan 430079, China

Received Date: 30 March 2017
Revised Date: 1 November 2017

Fund Project: This work was supported by the National Natural Science Foundation of China (No. 21577045).

In the present study, 2-methyl-4-(trifluoromethyl)thiazole-5-carboxylic acid (MTCA) was acted as hapten of thifluzamide to synthesize artificial antigens. Immunogen MTCA-bovine serum albumin (BSA) was synthesized by active ester method. Meanwhile, three coating antigens, MTCA-OVA-1, MTCA-OVA-2 and MTCA-OVA-3 were synthesized by active ester method, mixed anhydride method and N,N-carbonyldiimidazole/4-dimethylaminopyridine (CDI/DMAP) method, respectively. Anti-thifluzamide polyclonal antibody with high specificity was obtained from the immunized mice, then indirect competitive enzyme linked immunosorbent assay (ic-ELISA) method for thifluzamide detection was developed by using MTCA-OVA-3 and polyclonal antibody. The linear detection range in ic-ELISA was 0.08-10 mg/L with an IC₅₀ of 1.39 mg/L and a LOD (IC₁₀) of 0.08 mg/L. The recoveries for spiked samples including tap water, lake water and wheat ranged from 72.0%-128.3% in ic-ELISA method. Good correlation (R²=0.9994) was obtained between the results of ic-ELISA and those of HPLC analysis. The proposed ic-ELSA was promising for rapid detection of thifluzamide in water and agricultural products.
- Thifluzamide,
- Artificial antigen,
- Polyclonal antibody,
- Enzyme linked immunosorbent assay
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