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.
Hardouin
Julie
J
Laboratory of Protein Biochemistry and Proteomics, UMR CNRS 7033, UFR SMBH, Paris13 University, Bobigny, France. hardouin@smbh.univ-paris13.fr
Joubert-Caron
Raymonde
R
Caron
Michel
M
D000595:Amino Acid Sequence; D001324:Autoantigens / Q000032:analysis; D015415:Biomarkers / Q000032:analysis*; D018938:Caco-2 Cells; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods; D006801:Humans; D013058:Mass Spectrometry / Q000295:instrumentation / Q000379:methods; D008422:Materials Testing; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods; D008969:Molecular Sequence Data; D011506:Proteins / Q000032:analysis*; D015203:Reproducibility of Results; D020539:Sequence Analysis, Protein
D001324:Autoantigens; D015415:Biomarkers; D011506:Proteins