TR&D 4

Chromatographic & Electrophoretic Separations Optimized for Native MS — PIs: Susan Olesik, Abraham Badu-Tawiah

Direct infusion of native protein samples into the mass spectrometer is problematic when the sample complexity is too high and causes ion suppression or signal overlap. We are improving methods for the separation of native/folded proteins and protein complexes using chromatographic and electrophoretic techniques, both offline and directly coupled online with MS analysis. In addition, improved ionization approaches for coupling the separation method to the mass spectrometer will optimize chance of remaining native.

 

Aim 1

Chromatography and offline separation (SEC and IEX development/DESI-MS and integrated spray)
  • Develop offline and online SEC and IEC coupled to native MS for smaller sample sizes
  • Conversion of ESI to nESI at column exit for fractionation and control of charge states via contained electrospray
  • Develop offline IEX separations via improved planar electrospun nanofiber plates
  • Ionization and detection of native proteins separated by nanofiber plates via DESI-MS and integrated nESI

Aim 2

Improve capillary electrophoresis for separation of native proteins/complexes
  • Adapt commercial capillaries for separation under native electrolyte conditions
  • Develop novel capillary coatings and fillings for improved separation of native proteins and protein complexes