In vitro digestibility of beta-casein and beta-lactoglobulin under simulated human gastric and duodenal conditions: A multi-laboratory evaluation

G. Mandalari, K. Adel-Patient, Vibeke Barkholt, C. Baro, L. Bennett, M. Bublin, S. Gaier, G. Graser, G.S. Ladics, D. Mierzejewska, E. Vassilopoulou, Y.M. Vissers, L. Zuidmeer, N.M. Rigby, L.J. Salt, M. Defernez, F. Mulholland, A.R. Mackie, M.S.J. Wickham, E.N.C. Mills

    Research output: Contribution to journalJournal articleResearchpeer-review

    Abstract

    Initially the resistance to digestion of two cow's milk allergens, beta-casein, and beta-lactoglobulin (beta-Lg), was compared using a "high-protease assay" and a "low-protease assay" in a single laboratory. The low-protease assay represents an alternative standardised protocol mimicking conditions found in the gastrointestinal tract. For the high-protease assay, both proteins were incubated with either pepsin or pancreatin and digestion monitored by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and reverse phase-high performance liquid chromatography. The low-protease assay involved gastroduodenal digestion in the presence or absence of phosphatidylcholine (PC). Both beta-casein and beta-Lg were susceptible to hydrolysis by pepsin and pancreatin in the high-protease assay. In contrast, the kinetics of beta-casein digestion in the low-protease assay were slower, beta-Lg being pepsin resistant. During duodenal digestion, beta-Lg was gradually degraded and addition of PC slowed digestion. Subsequently, the reproducibility of the low-protease assay was assessed in 12 independent laboratories by visual assessment of the gels and densitometric analysis: the inter- and intra-laboratory variability was affected by sampling and electrophoresis method employed. The low-protease assay was shown to be reproducible. Future studies will extend these findings using a broader panel of proteins.
    Original languageEnglish
    JournalRegulatory Toxicology and Pharmacology
    Volume55
    Issue number3
    Pages (from-to)372-381
    ISSN0273-2300
    DOIs
    Publication statusPublished - 2009

    Keywords

    • Allergy
    • beta-Casein
    • beta-Lactoglobulin
    • In vitro digestion
    • Physiological protocol

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