HPLC-chip-mass spectrometry for protein signature identifications

This work investigates the use of an HPLC-chip microfluidic device interfaced to an IT mass spectrometer to search for biomarker signatures. To that end, the identification of autoantigens is chosen as a model. It not only constitutes a proof of concept model but also the growing interest in autoantibodies and autoantigens as new markers of diseases provides a practical application at the same time. The peptides are separated by the HPLC-chip system allowing suitable resolution and reproducibility. The determination of two parameters that characterize a peptide sequence during LC-MS/MS analyses, retention time (RT) and m/z ratio, improves the identification of a number of peptides derived from protein digests. These findings illustrate that accurate RT measurement obtained in a microfluidic device is useful to obtain mass/retention time (MRT) pairs for a given peptide, which can contribute to the definition of biomarker signatures.