Synthesis of Thifluzamide Artificial Antigens and Establishment of Enzyme Linked Immunosorbent Assay Detection Method
10.11895/j.issn.0253-3820.170206
- VernacularTitle:噻呋酰胺人工抗原的合成及酶联免疫吸附分析方法的建立
- Author:
Xiao Xiao LI
1
;
Mao Shu DING
;
Xu YANG
;
Lin Jun YUAN
Author Information
1. 华中师范大学生命科学学院 湖北省遗传调控和整合生物学重点实验室
- Keywords:
ThifluZamide;
Artificial antigen;
Polyclonal antibody;
EnZyme linked immunosorbent assay
- From:
Chinese Journal of Analytical Chemistry
2018;46(1):27-32
- CountryChina
- Language:Chinese
-
Abstract:
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.
