title: Improved detection specificity for plasma proteins by targeting cysteine-containing peptides with photo-SRM.
abstract: Targeted mass spectrometry using selected reaction monitoring (SRM) has emerged as an alternative to immunoassays for protein quantification owing to faster development time and higher multiplexing capability. However, the SRM strategy is faced with the high complexity of peptide mixtures after trypsin digestion of whole plasma or the cellular proteome that most of the time causes contamination, irremediably, by interfering compounds in the transition channels monitored. This problem becomes increasingly acute when the targeted protein is present at a low concentration. In this work, the merit of laser-induced photo-dissociation in the visible region at 473 nm implemented in an hybrid quadrupole linear ion-trap mass spectrometer (photo-SRM) was evaluated for detection specificity of cysteine-containing peptides in a group of plasma proteins after tagging with a dabcyl chromophore. Compared with conventional SRM, photo-SRM chromatograms have improved detection specificity for most of peptides monitored. Comparison of the signals obtained for the best proteotypic peptides in SRM mode and those recorded by photo-SRM of cysteine-containing peptides for the same proteins reveals either increased (up to 10-fold) or similar signal to photo-SRM detection. Finally, photo-SRM has extended response linearity across a calibration plot obtained by diluting human plasma in rat plasma, down to the lowest concentrations. Hence, photo-SRM may advantageously complement conventional SRM in assay of proteins in complex biological matrices.
authors: Université Lyon, 43 Boulevard du 11 Novembre 1918, 69622 Lyon, France.
authors: Quentin
authors: Q
authors: Enjalbert
authors: Marion
authors: M
authors: Girod
authors: Romain
authors: R
authors: Simon
authors: Jérémy
authors: J
authors: Jeudy
authors: Fabien
authors: F
authors: Chirot
authors: Arnaud
authors: A
authors: Salvador
authors: Rodolphe
authors: R
authors: Antoine
authors: Philippe
authors: P
authors: Dugourd
authors: Jérôme
authors: J
authors: Lemoine
mesh_terms: D000818:Animals; D001798:Blood Proteins / Q000737:chemistry*; D003545:Cysteine / Q000032:analysis; D006801:Humans; D013058:Mass Spectrometry / Q000295:instrumentation / Q000379:methods*; D010455:Peptides / Q000097:blood / Q000737:chemistry*; D051381:Rats; D012680:Sensitivity and Specificity
chemical_list: D001798:Blood Proteins; D010455:Peptides; D003545:Cysteine