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Immunoassay, biosensors and other non-chromatographic methods for agrochemical analysis
Abstract
This site is a comprehensive 58 page book chapter in pdf format on nonchromatographic methods of analysis for agrochemical contaminants (residues). The process of preparing and testing immunoassay tests is also discussed. Both biological and physical-based methods are discussed. In the former, it presents a wide variety of methods including the use of PCR. In the latter, it includes spectroscopy and voltammetry.
... In capture enzyme-linked immunosorbent method, a capture antibody (Y) is passively adsorbed on a solid phase (Figure 2, Shan et al., 2002 with modification). The target protein contained in the sample and the enzyme-labeled reported antibody (Y-E) is added. ...
... After homogenization, the supernatants will be collected and stored at -20 o C until further use in the ELISA analysis. Fig. 2. Enzyme-linked immunosorbent assay (ELISA) (Shan et al., 2002 with modification). ...
... The immunoassays are widely used in clinical analysis. However, other applications of immunoassays of increasing importance have been observed in other areas, such as in environmental control (Michal et al., 2007; Rodriguez-Mozaz et al., 2005; Velasco-Garcia, 2003) and in the quality control of food (Nandakumar et al., 2008; Skottrup et al., 2008; Choi, 2005; Sadik et al., 2004; Gaag et al., 2003; Shan et al., 2002). The use of biosensors along the last 20 years has been an approach for immunoassays which has had important and interesting results (Skottrup et al., 2008; Wang et al., 1998; Bergveld, 1991). ...
... Another advantage of the CPE in comparison with other solid electrodes is its regeneration. The electrode surface can simply be regenerated, after some assays, by only removing the corroded or used layer and polishing the surface, since the volume of the paste serves as a protein reservoir (Shan et al., 2002). The behavior of CPE modified with peroxidase (HRP-CPE) using several kinds of commercial graphite powders and certain additive elements known to act as promoters or stabilizers has been investigated by many investigators. ...
This paper is an overview of the recent developments in immunosensors, which have attracted considerable attention. Immunosensors can play an important role in the improvement of public health by providing applications for which rapid detection, high sensitivity, and specificity are important, in areas such as clinical chemistry, food quality, and environmental monitoring. This review focuses on the current research in immunoassay methods based on electrochemical detection for the analysis of environmental samples or medical diagnostic methods with emphasis on recent advances, challenges and trends. Technological aspects in the development of immunosensors such as kinetics of biomolecular interaction, techniques of immobilization, simplification of assay procedures, immunointeration and catalytic studies and system miniaturization are presented.
... Because, of its sensitivity, ruggedness and inexpensive characteristics, it is a choice of breeders for testing of unapproved events, and determining GM content ensuring compliance with non-GM labeling requirements. [16][17][18][19][20][21] Various ELISA and immunochromatographic strip formats for detecting Cry1Ab, Cry1Ac and Cry2Ab are already present in the market. [22][23][24] However, there is a pressing need to develop and Sandwich ELISA. ...
Immunoassays are being used extensively for the identification of genetically modified crops. Keeping pace with development of newer transgene crops, detection tests are also formatted for such crops and products. A sandwitch ELISA test for quantification of Cry2Ab protein was developed. The assay employed polyclonal rabbit IgG as capture antibody and monoclonal mouse anti-Cry2Ab -IgG as detection antibody. Addition of polyvinylpyrrolidone (2%) and dithiothritol (1%) during protein extraction from transgene cotton seeds ensured minimal interference with protein assay. The test was repeatedly validated with cotton seed samples. The limit of detection was 1pg/g and limit of quantification 16 pg/g. The assay did not show any cross reactivity with other non-target GM proteins i.e. Cry1Ab, Cry1Ac, EPSPS and Vip. This assay is the most sensitive one for Cry2Ab detection than the currently available test.
... Because, of its sensitivity, ruggedness and inexpensive characteristics, it is a choice of breeders for testing of unapproved events, and determining GM content ensuring compliance with non-GM labeling requirements. [16][17][18][19][20][21] Various ELISA and immunochromatographic strip formats for detecting Cry1Ab, Cry1Ac and Cry2Ab are already present in the market. [22][23][24] However, there is a pressing need to develop and Sandwich ELISA. ...
Immunoassays' a sensitive technique has a wide application for detection of antigen, especially for the detection of growing genetically modified crops, which are underwent field trial. Here, we have developed for the quantitative detection of Cry2Ab protein expressed in GM crops. A Cry2Ab-rabbit-IgG act as capture antibody and Cry2Ab mouse monoclonal antibody behave as a detecting antibody, which is employed in sandwich ELISA. A 2% polyvinylpyrrollidone and 1% dithiothritol was utilized in protein extraction from cotton seed to avert interfering agents. The developed assay was validated with GM cotton samples, limit of detection was found 1 pg/ml and range to a limit of quantification 16 pg/ml. The developed immunoassay does not show any cross reactivity with other non target GM proteins like Cry1Ab, Cry1Ac, EPSPS, Vip.
The development of multianalyte immunoassays constitutes a main research issue in the field of bioanalytical techniques. In the present study, class-specific antibodies against the three members of the anilinopyrimidine family of fungicides (pyrimethanil, cyprodinil, and mepanipyrim) were raised by using a bioconjugate of a rationally designed hapten [5-(6-methyl-2-(phenylamino)pyrimidin-4-yl)pentanoic acid]. Highly sensitive immunoassays were developed for the generic determination of these compounds, using the competitive enzyme-linked immunosorbent assay (ELISA). Particularly, a direct antibody-coated competitive ELISA afforded identical sensitivity for the three anilinopyrimidines, with IC50 values of 0.26, 0.27, and 0.25 μg L⁻¹ for pyrimethanil, cyprodinil, and mepanipyrim, respectively. This immunoassay was fully characterized and applied to the multianalyte determination of anilinopyrimidine fungicides in white and red wines, with a limit of quantification of 1 μg L⁻¹, average recoveries from 93.1% to 114.4%, and relative standard deviations lower than 20%. Commercial wine samples were analyzed and those containing detectable anilinopyrimide residues were verified by a reference chromatographic technique.
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