Capillary isoelectric focusing-electrospray mass spectrometry for protein analysis

Journal Article · Sun Oct 01 04:00:00 EDT 1995 · Analytical Chemistry (Washington)

DOI:https://doi.org/10.1021/ac00115a021· OSTI ID:111219

Tang, Q; Harrata, A K; Lee, C S ^[1]

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1. Ames Lab., IA (United States)

On-line combination of capillary isoelectric focusing (CIEF) with electrospray mass spectrometry (ESMS) as a two-dimensional separation system is demonstrated. Mixtures of model proteins including cytochrome c (horse heart), myoglobin (horse heart), and carbonic anhydrase II (bovine erythrocyte) are focused and cathodically mobilized in a polyacrylamide-coated capillary. At the end of CIEF capillary, the mobilized protein zones are analyzed by mass spectrometry coupled on-line to an electrospray interface with a coaxial sheath flow configuration. The effects of carrier ampholyte concentration on the CIEF separation and the protein electrospray ionization mass spectra are presented and discussed. In this study, the focusing effect of CIEF permits analysis of very dilute protein samples. A typical concentration factor of 50-100 times is observed. The concentration detection limit of myoglobin for a full-scan CIEF-ESMS analysis is in the range of 10{sup -7} M, 2 orders of magnitude over that possible with normal capillary zone electrophoresis ESMS. 35 refs., 5 figs., 2 tabs.

• Sponsoring Organization: USDOE
• OSTI ID: 111219
• Journal Information: Analytical Chemistry (Washington), Journal Name: Analytical Chemistry (Washington) Journal Issue: 19 Vol. 67; ISSN ANCHAM; ISSN 0003-2700
• Country of Publication: United States
• Language: English

Related Subjects

40 CHEMISTRY
55 BIOLOGY AND MEDICINE
BASIC STUDIES
CAPILLARIES
CYTOCHROMES
FOCUSING
MASS SPECTROSCOPY
MYOGLOBIN
NUMERICAL DATA
PROTEINS
ULTRAVIOLET RADIATION