Development and Validation of an Enzyme-Linked Immunosorbent Assay (ELISA) for Quantification of Lysozyme in Cheese

A competitive enzyme-linked immunosorbent assay (ELISA) was developed to quantify the amount of the preservative and potential allergen lysozyme in cheese using a commercially available monoclonal antibody against hen egg white lysozyme. The limit of detection for lysozyme in a cheese matrix amounted to 2.73 ng/mL, and the working range comprises 3.125-800 ng/mL. Intra- and interassay coefficients of variation were lower than 12%. Neither cross-reactivity with alpha-lactalbumin and human lysozyme nor unspecific interference with matrix components was observed. The recovery of lysozyme-spiked cheese ranged from 87.4 to 93.6% at four concentrations (50, 100, 200, and 400 mg/kg). The ELISA method was also compared to a high-performance liquid chromatography (HPLC) method, confirming the reliability and accuracy of the ELISA. A total of 21 commercially available cheese samples produced with and without lysozyme were analyzed with ELISA as well as HPLC. Both methods showed good agreement with a correlation index of R2=0.990.