Enzyme-linked immunosorbent assay (ELISA) is a widely used analytical technique for food allergen detection and quantification. Validating ELISA protocols is important for both assay developers and end users as it ensures method reliability. This chapter describes the protocols for validating the sensitivity, specificity, precision, accuracy, robustness, and ruggedness of an ELISA. Example procedures are also provided for sample preparation, allergen extraction, and ELISA operation.
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