Enzyme-Linked Immunosorbent Assay by Image Analysis Using a Charge-Coupled Device Array Detector

• Published in: Analytical biochemistry 1996-07, Vol.239 (1), p.2-7
• Main Authors: Sánchez, F.Garcı́a, Dı́az, A.Navas, Lovillo, J.
• Format: Article
• Language: English
• Subjects: 2,4-Dichlorophenoxyacetic Acid - analogs & derivatives; 2,4-Dichlorophenoxyacetic Acid - analysis; Animals; Drug Residues; Enzyme-Linked Immunosorbent Assay - instrumentation; Enzyme-Linked Immunosorbent Assay - methods; Evaluation Studies as Topic; Fruit - chemistry; Image Processing, Computer-Assisted; Rabbits; Reference Standards; Sensitivity and Specificity; Spectrometry, Fluorescence
• ISSN: 0003-2697; 1096-0309
• DOI: 10.1006/abio.1996.0283

This paper describes a fluorescence enzyme-linked immunosorbent assay (ELISA) for the quantification of (±)-2-(2,4-dichlorophen-oxy)propionic methyl ester (dichlorprop methyl ester). Antibodies for dichlorprop methyl ester were produced by immunizing rabbits with a conjugate of dichlorprop methyl ester with bovine serum albumin. Data acquisition on microtiter wells is performed by a spectrofluorometer through a fiber optic and by a charge-coupled device camera. A correlation was obtained between the image analysis data on ELISA and the data acquired by the spectrofluorometer. The results demonstrate that the fluorescence image analysis performed by the charge-coupled device detector is applicable to ELISA, and the analysis time, sensitivity, and precision of the ELISA procedure are compared to conventional fluorescence ELISA performed by the spectrofluorometer. The ELISA procedure was selective for structurally similar compounds or usually found in formulation pesticides. Concentrations for 50% displacement curves were dichlorprop, 83.59 μg/ml, and 2,4,5-T, 388.23 μg/ml; triclopyr, ioxynil, bentazone, and MCPA had no response.