TY - JOUR
TI - In vitro digestibility of β-casein and β-lactoglobulin under simulated human gastric and duodenal conditions: A multi-laboratory evaluation
AU - Mandalari, G.
AU - Adel-Patient, K.
AU - Barkholt, V.
AU - Baro, C.
AU - Bennett, L.
AU - Bublin, M.
AU - Gaier, S.
AU - Graser, G.
AU - Ladics, G.S.
AU - Mierzejewska, D.
AU - Vassilopoulou, E.
AU - Vissers, Y.M.
AU - Zuidmeer, L.
AU - Rigby, N.M.
AU - Salt, L.J.
AU - Defernez, M.
AU - Mulholland, F.
AU - Mackie, A.R.
AU - Wickham, M.S.J.
AU - Mills, E.N.C.
JO - Regulatory Toxicology and Pharmacology
PY - 2009
VL - 55
TODO - 3
SP - 372-381
PB - 
SN - 0273-2300, 1096-0295
TODO - 10.1016/j.yrtph.2009.08.010
TODO - beta casein;  beta lactoglobulin;  pancreatin;  pepsin A;  phosphatidylcholine, allergy;  article;  controlled study;  densitometry;  digestion;  duodenum;  electrophoresis;  enzyme assay;  food safety;  in vitro study;  laboratory test;  nonhuman;  nucleotide sequence;  polyacrylamide gel electrophoresis;  priority journal;  protein hydrolysis;  reproducibility;  reversed phase high performance liquid chromatography;  simulation;  stomach, Allergens;  Animals;  Caseins;  Chromatography, High Pressure Liquid;  Digestion;  Duodenum;  Electrophoresis, Polyacrylamide Gel;  Humans;  Lactoglobulins;  Milk;  Pancreatin;  Pepsin A;  Reproducibility of Results;  Sodium Dodecyl Sulfate;  Stomach
TODO - Initially the resistance to digestion of two cow's milk allergens, β-casein, and β-lactoglobulin (β-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 β-casein and β-Lg were susceptible to hydrolysis by pepsin and pancreatin in the high-protease assay. In contrast, the kinetics of β-casein digestion in the low-protease assay were slower, β-Lg being pepsin resistant. During duodenal digestion, β-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. © 2009 Elsevier Inc.
ER -