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Binding properties of a monoclonal antibody against the Cry1Ab from Bacillus Thuringensis for the development of a capillary electrophoresis competitive immunoassay

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Abstract

Experimental work performed was aimed at the assessment of a competitive capillary electrophoresis immunoassay with laser-induced fluorescence (CEIA-LIF) detection for the determination of the Cry1Ab endotoxin from Bacillus thuringensis. The binding constant of a monoclonal antibody, raised against the insecticide protein Cry1Ab, was determined on a microplate by indirect enzyme-linked immunosorbent assay (ELISA) and compared with that obtained in-capillary under nonequilibrium separation conditions. The two binding constants appear comparable—(5.0 ± 1.2) × 106 M−1 and (9.06 ± 5.7) × 106 M−1—reflecting good preservation of the antibody binding behavior in the capillary electrophoresis format. These results allow use of a calibration curve possible between 0.2 and 150 nM of endotoxin protein, with a limit of detection of 0.5 nM (33 μg L−1). Preliminary recovery experiments on maize extracts spiked with known amounts of Cry1Ab endotoxin also showed promising results in detecting the toxin in complex real matrices.

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Acknowledgements

The authors wish to thank the Prof. Marzari of the University of Trieste (Italy) for kindly supplying the transgenic Cry1Ab and the monoclonal antibody against the toxin.

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Authors and Affiliations

  1. Department of Analytical Chemistry, University of Torino, 10125, Torino, Italy

    Cristina Giovannoli, Laura Anfossi, Claudio Baggiani & Gianfranco Giraudi

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  1. Cristina Giovannoli
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  2. Laura Anfossi
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  3. Claudio Baggiani
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  4. Gianfranco Giraudi
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Correspondence to Cristina Giovannoli.

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Giovannoli, C., Anfossi, L., Baggiani, C. et al. Binding properties of a monoclonal antibody against the Cry1Ab from Bacillus Thuringensis for the development of a capillary electrophoresis competitive immunoassay. Anal Bioanal Chem 392, 385–393 (2008). https://doi.org/10.1007/s00216-007-1811-0

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  • DOI: https://doi.org/10.1007/s00216-007-1811-0

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