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Science 30 August 1996:
Vol. 273. no. 5279, pp. 1199 - 1202
DOI: 10.1126/science.273.5279.1199
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Reports

Attomole Protein Characterization by Capillary Electrophoresis-Mass Spectrometry

Gary A. Valaskovic, Neil L. Kelleher, Fred W. McLafferty *

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 approx 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.

Department of Chemistry, Baker Laboratory, Cornell University, Ithaca, NY 14853, USA.
*   To whom correspondence should be addressed.


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