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

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† Department of Chemistry and Pharmacy, Food Chemistry, Emil Fischer Center, University of Erlangen-Nuremberg, Schuhstrasse 19, 91052 Erlangen, Germany
‡ Bavarian Health and Food Safety Authority, Eggenreuther Weg 43, 91058 Erlangen, Germany
*To whom correspondence should be addressed. Telephone: +49-9131-8524102. Fax: +49-9131-8522587. E-mail: [email protected]
Cite this: J. Agric. Food Chem. 2010, 58, 1, 76–81
Publication Date (Web):November 17, 2009
https://doi.org/10.1021/jf9025019
Copyright © 2009 American Chemical Society
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Abstract

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 α-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.

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