A highly sensitive immunoassay for atrazine based on covalently linking the small molecule hapten to a urea–glutaraldehyde network on a polystyrene surface
| Autor: | Wenjing Sun, Na Sai, Zhong Sun, Yuntang Wu, Guowei Huang, Guanggui Yu |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
Immunology
Enzyme-Linked Immunosorbent Assay 010501 environmental sciences Sensitivity and Specificity Zea mays 01 natural sciences chemistry.chemical_compound Microtiter plate Reference Values medicine Humans Urea Immunology and Allergy Sulfhydryl Compounds Atrazine 0105 earth and related environmental sciences Pharmacology Detection limit Chromatography medicine.diagnostic_test Herbicides 010401 analytical chemistry Water Oryza Small molecule 0104 chemical sciences chemistry Glutaral Covalent bond Immunoassay Polystyrenes Environmental Pollutants Glutaraldehyde Haptens Hapten |
| Zdroj: | International Immunopharmacology. 40:480-486 |
| ISSN: | 1567-5769 |
| DOI: | 10.1016/j.intimp.2016.10.003 |
| Popis: | A new enzyme-linked immunosorbent assay (ELISA) for atrazine was developed based on covalent bonding of the small molecule hapten, 2-mercaptopropionic acid-4-ethylamino-6-isopropylamino-1,3,5-triazine (MPA–atrazine), to urea-glutaraldehyde (UGA)-treated microtiter plates. In this assay, the microtiter plate surface was treated with the UGA network to both introduce amino groups, which were used to cross-link with the hapten carboxylate groups, and efficiently prevent non-specific adsorption of antibodies, which successfully eliminated the time-consuming routine blocking step. Compared with HNO3-H2SO4-APTES-hapten coated ELISA (modified with a HNO3-H2SO4-APTES mixture and covalent-linked hapten) and conventional ELISA (coated with hapten–carrier protein conjugates), the novel ELISA format increased the sensitivity by approximately 3.5-fold and 7.5-fold, respectively, and saved 2.5 h and 34 h of coating hapten time, respectively. The method's 50% inhibition concentration for atrazine was 5.54 ng mL− 1, and the limit of detection was 0.16 ng mL− 1 after optimization of reaction conditions. Furthermore, the ELISA was adapted for analysis of atrazine in corn, rice, and water samples, demonstrating recoveries of 90%–108%. Thus, the assay provides a convenient alternative to conventional, laborious immunoassays for routine supervision of residue detection in food and the environment. |
| Databáze: | OpenAIRE |
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