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Published on: March 6, 2019
Exploring alternative hapten tethering sites for high-affinity anti-picoxystrobin antibody generation
Javier Parra1, Francesc A Esteve-Turrillas, Antonio Abad-Somovilla
1Department of Biotechnology, Instituto de Agroquímica y Tecnología de Alimentos-Consejo Superior de Investigaciones Científicas, 46980 Paterna, València, Spain.
Analytical Biochemistry
|June 10, 2011
Summary
Antibody affinity for the fungicide picoxystrobin depends on hapten linker site. Site heterology in immunoassays improved detection limits for fungicide residue analysis in soybean sprouts.
Area of Science:
- Agrochemicals
- Immunochemistry
- Analytical Chemistry
Background:
- Hapten modification is crucial for antibody generation and immunoassay sensitivity.
- The linker tethering site's impact on antibody performance requires further investigation for pesticide residue analysis.
Purpose of the Study:
- To synthesize picoxystrobin derivatives with varying linker positions.
- To generate and characterize polyclonal antibodies against picoxystrobin.
- To develop and optimize a sensitive immunoassay for picoxystrobin detection.
Main Methods:
- Synthesis of three picoxystrobin hapten derivatives with different linker positions.
- Production and characterization of polyclonal antibodies using protein conjugates.
- Development and optimization of competitive enzyme-linked immunosorbent assays (ELISAs).
Main Results:
- All synthesized haptens generated highly specific antibodies.
- Significant differences in antibody affinity for free picoxystrobin were observed based on hapten structure.
- An indirect heterologous immunoassay achieved a limit of detection of 0.02 μg/L for picoxystrobin.
- The method provided a practical limit of quantification of 5 μg/kg in soybean sprouts.
Conclusions:
- The linker tethering site in haptens significantly influences antibody affinity and immunoassay performance.
- Site heterology is an effective strategy for enhancing detectability in pesticide residue analysis.
- The developed immunoassay is suitable for monitoring picoxystrobin levels in food commodities.
