Premium
Analysis of human blood serum using the off‐line coupling of capillary isoelectric focusing to matrix‐assisted laser desorption/ionization time of flight mass spectrometry
Author(s) -
Crowley Timothy A.,
Hayes Mark A.
Publication year - 2005
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200401212
Subject(s) - isoelectric focusing , chromatography , chemistry , mass spectrometry , analytical chemistry (journal) , capillary action , ionization , matrix (chemical analysis) , desorption , elution , sample preparation , matrix assisted laser desorption/ionization , capillary electrophoresis , materials science , adsorption , ion , biochemistry , organic chemistry , composite material , enzyme
Off‐line coupling of capillary IEF (CIEF) with matrix‐assisted laser desorption/ionization mass spectrometry was utilized for the analysis of human blood serum. Serum proteins were initially separated by CIEF, and fractions of the isoelectric separation were eluted sequentially to a MALDI‐TOF MS sample target. During pressure elution of the CIEF sample, voltage was maintained across the capillary system utilizing a sheath flow arrangement to minimize band broadening induced by the laminar flow field. Both p I and mass information were obtained from the complex biological sample, similar to traditional 2‐DE techniques, and the platform was faster (hours versus days), more automatable, and simpler than 2‐DE. The volume of raw sample present in the actual analysis was approximately 100 nL, making this technique well suited for very rare specimens. Additionally, the speed and simplicity of the technology make it an attractive technique for performing initial comparative analyses of complex samples.