Heart-cut nano-LC-CZE-MS for the characterization of proteins on the intact level.
Electrophoresis
; 40(7): 1061-1065, 2019 04.
Article
en En
| MEDLINE
| ID: mdl-30575976
ABSTRACT
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.
Palabras clave
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Ribonucleasas
Idioma:
En
Revista:
Electrophoresis
Año:
2019
Tipo del documento:
Article
País de afiliación:
Alemania