Towards the Analysis of High Molecular Weight Proteins and Protein complexes using TIMS-MS.
In the present work, we demonstrate the potential and versatility of TIMS for the analysis of proteins, DNA-protein complexes and protein-protein complexes in their native and denatured states. In addition, we show that accurate CCS measurement are possible and in good agreement with previously reported CCS values using other IMS analyzers (<5% difference). The main challenges for the analysis of high mass proteins and protein complexes in the mobility andm/zdomain are described. That is, the analysis of high molecular weight systems in their native state may require the use of higher electric fields or a compromise in the TIMS mobility resolution by reducing the bath gas velocity in order to effectively trap at lower electric fields. This is the first report of CCS measurements of high molecular weight biomolecules and biomolecular complexes (~ 150 kDa) using TIMS-MS.
Department of Chemistry & Biochemistry, Florida International University, Miami, FL 33199, USA.
Paolo
P
Benigni
Department of Chemistry & Biochemistry, Florida International University, Miami, FL 33199, USA.
Rebecca
R
Marin
Department of Chemistry & Biochemistry, Florida International University, Miami, FL 33199, USA.
Juan Camilo
JC
Molano-Arevalo
Department of Chemistry & Biochemistry, Florida International University, Miami, FL 33199, USA.
Alyssa
A
Garabedian
Bruker Daltonics, Inc., Billerica, Massachusetts 01821, USA.
Jeremy J
JJ
Wolff
Bruker Daltonics, Inc., Billerica, Massachusetts 01821, USA.
Mark E
ME
Ridgeway
Bruker Daltonics, Inc., Billerica, Massachusetts 01821, USA.
Melvin A
MA
Park
Department of Chemistry & Biochemistry, Florida International University, Miami, FL 33199, USA; Biomolecular Science Institute, Florida International University, Miami, FL 33199, USA.
Francisco
F
Fernandez-Lima
IMS-MS; Ion mobility spectrometry – Mass Spectrometry; TIMS; ion—neutral collisional cross sections; proteins