A Label-free, Data-independent Approach to Defining the Gluten Proteome

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Sophie Bromilow, Lee A. Gethings, Peter Shewry, E.N. Clare Mills
Waters, University of Manchester, Rothamsted Research
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This study highlights improved gluten extraction efficiency using the acid labile detergent RapiGest followed by identification of the celiac-toxic motif regions using a label-free LC-HDMSE (LC-DIA-IM-MS) approach.

The extraction efficiency of gluten proteins has shown to improve with buffers containing RapiGest when compared with other extraction buffers. Subsequent analysis with SYNAPT G2-Si provided 25% more protein identifications when using HDMSE compared with a competitor platform, which utilized a DDA strategy. The number of unique peptides identified was also 10-fold higher for HDMSE data, providing a greater number of glutenin and associated celiac-toxic motifs. Qualitative and quantitative information is obtained within a single acquisition. Label-free quantitation varies depending on the sub-group of gluten proteins.

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