Rapid characterization of structure-dependency gas-phase ion/ion reaction via accumulative tandem MS.

To enable the rapid detection of biomolecule reactivity and reaction sites, we developed a method based on gas-phase ion/ion reaction and accumulative tandem mass spectrometry (MS). Structure-dependency reactions in gas-phase were performed between biomolecule ions and their reaction partner ions with opposite polarities in a quadrupole ion trap. Gas-phase peptide bioconjugation with pyridoxal-5-phosphate (PLP) was chosen as a proof-of-principle example. It is found that the Coulomb attraction force holds reaction partners close together, which increasing the reaction probability. Post reaction, reaction sites were identified by the consequent accumulative tandem MS method, in which informative product ions in low abundance were enriched by more than 100 times in another quadrupole ion trap. With enough product ions, tandem MS was performed, and reaction sites could be identified unambiguously. Since those reactions are normally biomolecular structure dependent, density functional theory (DFT) calculations were also carried out to understand the reaction mechanism. The method allows for rapid characterization of structure dependent reactivity of a biomolecule, and opens a new avenue for drug development and biomolecule structure analyses.