Apparent discrepancies between immunochemical methods used for ovalbumin recognition in food
Article first published online: 19 SEP 2006
Copyright © 1989 John Wiley & Sons, Ltd
Journal of the Science of Food and Agriculture
Volume 47, Issue 3, pages 311–325, 1989
How to Cite
Breton, C., Thanh, L. P., Dubray, G. and Paraf, A. (1989), Apparent discrepancies between immunochemical methods used for ovalbumin recognition in food. J. Sci. Food Agric., 47: 311–325. doi: 10.1002/jsfa.2740470306
- Issue published online: 19 SEP 2006
- Article first published online: 19 SEP 2006
- Manuscript Accepted: 21 JUL 1988
- Manuscript Received: 8 JUN 1988
- European Economic Communication. Grant Number: 4179/A
- Association pour la Reserche contre le Cancer. Grant Number: 9177/B
- liquid-phase precipitation
The specificity of rabbit anti-native ovalbumin antibodies against native ovalbumin (NOA) or heat-denatured ovalbumin (HDOA) was studied by affinity chromatography, ELISA techniques, quantitative immuno-precipitation and HPLC. Affinity-purified antibodies were prepared either by passing antisera successively on an NOA column and an HDOA column, or vice versa. Antisera were found to contain about 85 % of antibodies directed against NOA, 7% specific for HDOA and 8% retained by both columns.
It is shown that antibodies specific for NOA fully precipitated NOA in liquid-phase precipitation but were unable to bind NOA in ELISA using NOA-coated plates, while recognising NOA trapped on the solid phase by a first layer of antibodies: ovalbumin was thus completely denatured during interaction with plastic as shown also by the fact that antibodies specific for HDOA reacted with NOA-coated plates.
These results point out the need to modify ELISA tests for monoclonal antibodies selection and for structural studies of food molecules in complex mixtures. However, this observation, which holds true for ovalbumin, might not apply to all antigens.