TY  - JOUR
U1  - Zeitschriftenartikel, wissenschaftlich - begutachtet (reviewed)
A1  - Jooß, Kevin
A1  - Scholz, Nico
A1  - Meixner, Jens
A1  - Neusüß, Christian
T1  - Heart-cut nano-LC-CZE-MS for the characterization of proteins on the intact level
JF  - Electrophoresis
N2  - Multidimensional separation techniques play an increasingly important role in separation science, especially for the analysis of complex samples such as proteins. The combination of reversed-phase liquid chromatography in the nanoscale and CZE is especially beneficial due to their nearly orthogonal separation mechanism and well-suited geometries/dimensions. Here, a heart-cut nano-LC-CZE-MS setup was developed utilizing for the first time a mechanical 4-port valve as LC-CE interface. A model protein mixture containing four different protein species was first separated by nano LC followed by a heart-cut transfer of individual LC peaks and subsequent CZE-MS analysis. In the CZE dimension, various glycoforms of one protein species were separated. Improved separation capabilities were achieved compared to the 1D methods, which was exemplarily shown for ribonuclease B and its different glycosylated forms. LODs in the lower μg/mL range were determined, which are considerably lower compared to traditional CZE-MS. In addition, this study represents the first application of an LC-CE-MS system for intact protein analysis. The nano-LC-CZE-MS system is expected to be applicable to various other analytical challenges.
KW  - Chromatography, Reverse-Phase/instrumentation/methods
KW  - Electrophoresis, Capillary/instrumentation/methods
KW  - Glycosylation
KW  - Hydrogen-Ion Concentration
KW  - Limit of Detection
KW  - Ribonucleases/analysis
KW  - Tandem Mass Spectrometry/instrumentation/methods
Y1  - 2019
SN  - 0173-0835
SS  - 0173-0835
U6  - https://doi.org/10.1002/elps.201800411
DO  - https://doi.org/10.1002/elps.201800411
PM  - 30575976
VL  - 40
SP  - 1061
EP  - 1065
ER  -