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  • A targeted mass spectrometry method for the accurate label-free quantification of immunogenic gluten peptides produced during simulated digestion of food matrices.

A targeted mass spectrometry method for the accurate label-free quantification of immunogenic gluten peptides produced during simulated digestion of food matrices.

MethodsX (2020-10-10)
Olivia Ogilvie, Nigel Larsen, Kevin Sutton, Laura Domigan, Juliet Gerrard, Nicholas Demarais, Sarah Roberts
ABSTRACT

Mass spectrometry (MS) is an emerging method to determine the accurate concentration of immunogenic gluten peptides. It is of interest to quantify specific peptides within the gluten peptidome due to the role they play in the activation of the celiac immune cascade. Celiac disease is an autoimmune disorder triggered in genetically susceptible individuals by the presence of specific gluten peptides that resist digestion in the gastrointestinal tract. The protocol detailed within this paper can accurately quantify (label-free) the concentration of six immunogenic gluten peptides (including the 33mer) released from a food matrix using the INFOGEST in vitro digestion protocol. This method can be used to monitor small changes in the concentration of these marker peptides in response to exogenous factors such as plant-breeding, fermentation or food processing.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Pepsin from porcine gastric mucosa, lyophilized powder, ≥2,500 units/mg protein (E1%/280)
Sigma-Aldrich
α-Amylase from porcine pancreas, PMSF Treated, Type I-A, saline suspension, ≥1000 units/mg protein (E1%/280)
Sigma-Aldrich
α-Chymotrypsin from bovine pancreas, Type II, lyophilized powder, ≥40 units/mg protein
Sigma-Aldrich
Trypsin from porcine pancreas, Type IX-S, lyophilized powder, 13,000-20,000 BAEE units/mg protein