Capillary Electrophoresis

Description: Sciex CE instruments are normally utilized for the separation of denatured protein samples. Some of the first experiments that we performed were the separation of denatured standard samples including intact antibody proteoforms. A close collaboration with Sciex has delivered support to make quick progress on this front. Significant progress has been made on the use of CE for the separation of native proteins and their complexes. Publications have encompassed CE standard operating procedure guides to perform native sample preparation, method development, separations, troubleshooting tips, and CE top-down fragmentation analysis to provide feasible scientific community adoptability for this CE technology we are developing. Additional works feature applicability of CE development into the applications realm including characterization of synthetic and endogenous nucleosomes.

 

Demonstration of standard native proteoform separation and analysis by CZE-Top-Down MS.

 

TR&D 5 Development Status: On going

Publications:

1.Jooß, K., McGee, J.P., Melani, R.D. and Kelleher, N.L. (2021), Standard procedures for native CZE-MS of proteins and protein complexes up to 800 kDa. ELECTROPHORESIS, 42: 1050-1059.

2.Kevin Jooß, Luis F. Schachner, Rachel Watson, Zachary B. Gillespie, Sarah A. Howard, Marcus A. Cheek, Matthew J. Meiners, Amin Sobh, Jonathan D. Licht, Michael-Christopher Keogh, and Neil L. Kelleher Separation and Characterization of Endogenous Nucleosomes by Native Capillary Zone Electrophoresis–Top-Down Mass Spectrometry. Anal. Chem. 93, 5151-5160 (2021)

3.Schachner, L.F., Jooß, K., Morgan, M.A. et al. Decoding the protein composition of whole nucleosomes with Nuc-MS. Nat Methods 18, 303–308 (2021)

Scientific Presentations:

ASMS 2021, ASMS 2020