Electron transfer dissociation for biopolymer sequence mass spectrometric analysis
AssigneeUniversity of Virginia Patent Foundation
InventorsDonald F. Hunt, Joshua J. Coon, John Edward Philip Syka, Jarrod A. Marto
Life of this patent
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Abstract
The present invention relates to a new method for fragmenting ions in a mass spectrometer through the use of electron transfer dissociation, and for performing sequence analysis of peptides and proteins by mass spectrometry. In the case of peptides, the invention promotes fragmentation along the peptide backbone and makes it possible to deduce the amino acid sequence of the sample, including modified amino acid residues, through the use of an RF field device.
Claims35
A method of mass spectrometry analysis based on electron transfer dissociation (ETD) of multiply charged organic and/or biomolecular cations, said method comprising the steps of
- introducing said multiply charged cations into an RF electric field ion containment device of a mass spectrometer;
- introducing polyaromatic hydrocarbon anions as gas-phase electron transfer reagents into said ion containment device;
- mixing the introduced polyaromatic hydrocarbon anions, or derivative anions thereof, and the multiply charged cations, or derivative multiply charged cations thereof, wherein said derivative anions and said derivative multiply charged cations are generated within said ion containment device during performance of said method, so as to facilitate electron transfer from the polyaromatic hydrocarbon anions, or said derivative anions thereof, to the multiply charged cations, or derivative multiply charged cations thereof, to induce cleavage of covalent bonds and produce fragment and/or dissociation product cations;
- and
- mass (m/z) analyzing and detecting said fragment and/or dissociation product cations or cations derived from said fragment and/or dissociation product cations for mass spectrometric analysis.
A method for analyzing the amino acid sequence of a polypeptide, said method comprising
- introducing multiply charged polypeptide cations into an RF ion trap;
- introducing polyaromatic hydrocarbon anions as gas-phase electron transfer reagents into an RF ion trap;
- mixing said polyaromatic hydrocarbon anions and multiply charged polypeptide cations so as to facilitate electron transfer from the polyaromatic hydrocarbon anions to the multiply charged polypeptide cations, thus inducing cleavage of covalent bonds and the production of electron transfer fragment and/or dissociation product cations;
- terminating the reactions by physically separating the remaining polyaromatic hydrocarbon anions from the electron transfer fragment and/or dissociation product cations;
- conducting m/z analysis of cations remaining in the trap.
A method for dissociating a polypeptide by negative electron transfer dissociation (NETD) for mass spectrometry analysis, said method comprising
- introducing multiply charged polypeptide anions into an RF electric field ion containment device;
- introducing gas-phase cations of an inert gas, into an RF electric field ion containment device;
- mixing the gas-phase inert gas cations and the polypeptide anions so as to facilitate electron transfer from the anions to the cations, and thus inducing the cleavage of covalent bonds and production of negative electron transfer fragment and/or dissociation product anions;
- and
- mass (m/z) analyzing and detecting said negative electron transfer dissociation product ions or ions derived from negative electron transfer dissociation product ions.
Dependencies are inferred from the claim text.