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Development of Two Immunoassay Formats To Detect β-Lactoglobulin: Influence of Heat Treatment on β-Lactoglobulin Immunoreactivity and Assay Applicability in Processed Food

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Milk proteins are commonly used as ingredients in the food industry because of their functional properties, but they can cause severe reactions in milk-allergic individuals. In this work, two enzyme-linked immunosorbent assay (ELISA) formats were developed to detect bovine β-lactoglobulin. The indirect competitive ELISA involved the use of anti–β-lactoglobulin antisera, and the sandwich ELISA involved the use of specific antibodies isolated using a β-lactoglobulin immunosorbent material. The effect of heat treatment on immunoreactivity of the protein in buffer and in milk was determined with both assays. The amount of immunoreactive protein in buffer and in milk decreased as determined by the sandwich ELISA, whereas the amount increased when measuring with the competitive ELISA. Several food products, including meat, bakery products, sauces, and snacks, were analyzed. With both assays, 10 of 11 products in which the ingredient list included the terms "powdered milk" or "milk proteins" contained β-lactoglobulin. However, the β-lactoglobulin concentration in these products obtained with the competitive ELISA were much higher than those obtained with the sandwich ELISA. These differences could be explained by the fact that the determination of β-lactoglobulin concentration by immunoassay is influenced by differences in antibody recognition of the protein present in highly processed foods. Therefore, the antigen-binding properties of antibodies used in a particular immunoassay are important for a correct interpretation of results obtained in food processed at high temperature.

Document Type: Research Article

Affiliations: Tecnología de los Alimentos, Facultad de Veterinaria, Miguel Servet 177, 50013 Zaragoza, Spain

Publication date: July 1, 2007

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    The Journal of Food Protection (JFP) is a refereed monthly publication. Each issue contains scientific research and authoritative review articles reporting on a variety of topics in food science pertaining to food safety and quality. The Journal is internationally recognized as the leading publication in the field of food microbiology with a readership exceeding 11,000 scientists from 70 countries. The Journal of Food Protection is indexed in Index Medicus, Current Contents, BIOSIS, PubMed, Medline, and many others.

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