on 06-18-202001:44 PM - edited on 10-15-202106:08 AM by AnalyteGuru
Mick Greer1, Daniel Lopez-Ferrer2, Romain Huguet2, Peter Verhaert3, Greg Foster4, Vlad Zabrouskov2, Andreas Huhmer2, Peter Raus5, Ken Durbin6, Joe Greer6, Ryan Fellers6, Rich Leduc6 ASMS 2020 Purpose: Apply newly developed Top-down Label Free Quantitation (LFQ).to measure proteoform level changes between age groups from human tear fluid.
Methods: Tear fluid was collected using simple non-invasive Schirmerstest strips. Samples were then submitted to Top-down LC/MS analysis with (Ultraviolet Photodissociationat 213 nm) UVPD enabled fragmentation. Resulting data was analyzed via ProSightPD4.0 (beta) for determination of quantitative changes in human tear proteoforms associated with patient age.
Results: Due to numerous crosslinked species observed in tear fluid, UVPD was selected for fragmentation of these proteoforms. A significant portion of the identified proteoforms were truncated species not known in UniProt, therefore Top-down MS with ProSightPDSubsequence analysis is the preferred method to accurately identify these species.