Innovative Instrumentation and Methods for the Identification of Intact Proteins in Mixtures and for Sequence Analysis of Antibodies and Posttranslationally-Modified, Intact Proteins on a Chromatograpic Time-Scale
University of Virgina
This lecture will focus on data generated with a new ion source that facilitates simultaneous generation of positively charged sample ions by electrospray ionization and negatively charged reagent ions for both electron transfer dissociation (ETD) and ion-ion proton transfer (IIPT) reactions on Orbitrap mass spectrometers. Implementation of multiple C-trap fills for enhanced sensitivity will be discussed and both parallel peak parking, and ion ejection strategies to facilitate protein separation and enhanced sequence coverage of intact proteins will be described. Use of IIPT/ETD facilitates near complete sequence coverage on many intact proteins and is ideally suited for locating multiple posttranslational modifications on the same protein molecule. Sequence analysis of antibodies with an enzyme reactor that generates 3-10 KDa fragments in seconds will also be discussed.
Presented at MSACL 2016 US