Validation of an enzyme-linked immunosorbent assay by using the Total Error Concept, the Accuracy Profiles and the Accuracy Index as alternatives to the traditional ICH approach

Abstract

The use of Total Error Concept for validating analytical methods is an approach that incorporates the sum of the Trueness and the Precision. This method also uses the accuracy profiles based on the tolerance intervals for whether a calibration model will give the quality results and it enables to control the risk in accepting an inappropriate methodology. The Total Error Concept, the Accuracy Profiles and the Accuracy Index were applied in this study for validating an enzyme-linked immunosorbent assay (ELISA) to determine the residual contents of ovalbumin in vaccines. This study tested a pure vaccine sample and spiked with three different concentrations (low, medium and high). Also an approach was used for evaluating the calculations methods which would yield the most accurate results. The obtained results in the concentrations range tested with the chosen model were: Trueness - ER% from 1.61 % to 12.15 %; intermediate precision - CV% from 6.91 % to 9.31 %; and the Total Error from 9.83 % to 19.07 %, which are in conformity with the international guides. This study showed that the method is reliable to evaluate the ovalbumin contents, and that the Total Error Concept approach has applicability for validating enzyme immunoassays.