Development of an immunoassay for determination of 2,4-dichlorophenoxyacetic acid (2,4-D) based upon the recombinant Fab fragment of 2,4-D specific antibody

Van Cuong Nguyen; Thi Dieu Thuy Nguyen; Hung Anh Dau; Thu Nga Tham; Dinh Thih Quyen; Till Bachmman; Rolf D. Schmid

doi:10.1117/12.440159

14 September 2001 Development of an immunoassay for determination of 2,4-dichlorophenoxyacetic acid (2,4-D) based upon the recombinant Fab fragment of 2,4-D specific antibody

Van Cuong Nguyen, Thi Dieu Thuy Nguyen, Hung Anh Dau, Thu Nga Tham, Dinh Thih Quyen, Till Bachmman, Rolf D. Schmid

Author Affiliations +

Proceedings Volume 4414, International Conference on Sensor Technology (ISTC 2001); (2001) https://doi.org/10.1117/12.440159
Event: International Conference on Sensing units and Sensor Technology, 2001, Wuhan, China

Abstract

To develop an immunoassay and further an immunosensor for 2,4-D based upon recombinant antibody, the Fab fragments of 2,4-D specific antibody were expressed in E. coli. Western blotting analysis of the periplasmic cell fractions shown that under the non-reducing condition only a single protein band at a molecular mass of 45-kDa, corresponding to the whole Fab fragment was detected. Antigen binding activity for 2,4-D was found only in the extract of cells bearing the 2,4-D plasmid. An immunoassay based on the competitive reaction of 2,4-D and enzyme tracer with 2,4-D Fab fragments immobilized on micro titer plates via rabbit anti-mouse IgC was developed. Using this assay, 2,4-D could be detected at concentration range of 0.5 (mu) g/1 to 10(mu) g/1. The center point of the 2,4-D test was found at a concentration of 5 (mu) g/l. The assay was applied for detection of 2,4-D in spiked orange samples, resulting in recovery rate of 90 percent. The immunoassay could be applied to monitor human exposure to 2,4-D from contamination in fruit samples.

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Van Cuong Nguyen, Thi Dieu Thuy Nguyen, Hung Anh Dau, Thu Nga Tham, Dinh Thih Quyen, Till Bachmman, and Rolf D. Schmid "Development of an immunoassay for determination of 2,4-dichlorophenoxyacetic acid (2,4-D) based upon the recombinant Fab fragment of 2,4-D specific antibody", Proc. SPIE 4414, International Conference on Sensor Technology (ISTC 2001), (14 September 2001); https://doi.org/10.1117/12.440159

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