Improved particle-packed HPLC/MS microchips for proteomic analysis.
J Sep Sci
; 33(21): 3283-91, 2010 Nov.
Article
en En
| MEDLINE
| ID: mdl-21049518
ABSTRACT
The influence of packing process parameters (packing pressure, application of ultrasound) and the stationary phase particle size (3.5 and 5 µm) on the chromatographic performance of HPLC/MS chips was systematically investigated for proteomic samples. First, reproducibility and detection limits of the separation were evaluated with a low-complexity sample of tryptic BSA peptides. The influence of adsorbent packing quality on protein identification was then tested with a typical proteomics sample of high complexity, a human plasma protein fraction (Cohn fraction IV-4). All HPLC/MS chips provided highly reproducible separations of these proteomic samples, but improved packing conditions and smaller particle sizes resulted in chromatograms with narrower peaks and correspondingly higher signal intensities. Improved separation performance increased the peak capacity, the number of identified peptides, and thus the sequence coverage in the proteomic samples, particularly for low sample amounts.
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Cromatografía Líquida de Alta Presión
/
Proteómica
/
Procedimientos Analíticos en Microchip
/
Espectrometría de Masas en Tándem
Tipo de estudio:
Evaluation_studies
Límite:
Humans
Idioma:
En
Revista:
J Sep Sci
Año:
2010
Tipo del documento:
Article
País de afiliación:
Alemania