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Hook effect

The high dose hook effect refers to measured levels of antigen displaying a significantly lower absorbance than the actual level present in a sample.

This appears when a simultaneous ELISA assay is saturated by a very high concentration of sample antigen binding to all available sites on both the solid phase antibody as well as the detection antibody and thereby preventing the sandwich-formation. The antigen-saturated detection antibodies in solution will be washed off giving a falsely low signal. A “hook” is observed in the curve when data is plotted as a signal versus antigen concentration.

To learn more about the ELISA procedure watch this short video: