Abstract
Electrospray ionization with an ultralow flow rate (≤4 nanoliters per minute) was used to directly couple capillary electrophoresis with tandem mass spectrometry for the analysis and identification of biomolecules in mixtures. A Fourier transform mass spectrometer provided full spectra (>30 kilodaltons) at a resolving power of ≈60,000 for injections of 0.7 × 10 −18 to 3 × 10 −18 mole of 8- to 29-kilodalton proteins with errors of <1 dalton in molecular mass. Using a crude isolate from human blood, a value of 28,780.6 daltons (calculated, 28,780.4 daltons) was measured for carbonic anhydrase, representing 1 percent by weight of the protein in a single red blood cell. Dissociation of molecular ions from 9 × 10 −18 mole of carbonic anhydrase gave nine sequence-specific fragment ions, more data than required for unique retrieval of this enzyme from the protein database.
Keywords
ChemistryMass spectrometryTandem mass spectrometryMolecular massCarbonic anhydraseCarbonic anhydrase IIElectrospray ionizationCapillary electrophoresisChromatographyElectrosprayAnalytical Chemistry (journal)EnzymeBiochemistry
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Publication Info
- Year
- 1996
- Type
- article
- Volume
- 273
- Issue
- 5279
- Pages
- 1199-1202
- Citations
- 391
- Access
- Closed
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Cite This
Gary A. Valaskovic,
Neil L. Kelleher,
Fred W. McLafferty
(1996).
Attomole Protein Characterization by Capillary Electrophoresis-Mass Spectrometry.
Science
, 273
(5279)
, 1199-1202.
https://doi.org/10.1126/science.273.5279.1199
Identifiers
- DOI
- 10.1126/science.273.5279.1199