Methods and Systems for LC-MS/MS Proteomic Genotyping
AssigneeLaboratory Corporation of America Holdings
InventorsChristopher Michael Shuford, Russell Philip Grant, Meghan Norris Bradley, Patricia Louise Miller Holland, Michael Levandoski
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Abstract
Disclosed are methods and systems using liquid chromatography/tandem mass spectrometry (LC-MS/MS and 2D-LC-MS/MS) for the proteomic analysis of genotypes. In certain embodiments, samples used in the analysis comprise dried bodily fluids.
Claims20
A method for determining a genotype of a gene of interest in a subject, the method comprising:
- providing a dried plasma sample containing a protein derived from the gene of interest, wherein the dried plasma sample is deposited on a solid substrate and comprises dried plasma separated from red blood cells;
- digesting the dried plasma sample to generate at least one allele specific surrogate peptide for the protein, thereby generating a digested sample;
- using mass spectrometry, detecting the at least one allele specific surrogate peptide in the digested sample;
- and
- determining the genotype of the subject based on results of the detecting the at least one allele specific surrogate peptide.
A method for determining a genotype of a subject, the method comprising:
- providing a dried plasma sample, wherein the dried plasma sample is deposited on a solid substrate and comprises dried plasma obtained from the subject and separated from red blood cells;
- digesting the dried plasma sample to generate a plurality of allele specific surrogate peptides for protein variants contained in the dried plasma, thereby generating a digested sample;
- using mass spectrometry, detecting the plurality of the allele specific surrogate peptides in the digested sample to generate a proteomic profile of the allele specific surrogate peptides;
- and
- determining the genotype of the subject based on the proteomic profile.
A system for determining genotype of a gene of interest in a subject, the system comprising:
- a station for subjecting a dried plasma sample to digestion to generate at least one allele specific surrogate peptide for a protein derived from the gene of interest, wherein the dried plasma sample is deposited on a solid substrate and comprises dried plasma separated from red blood cells;
- optionally, a station for chromatographic purification of the at least one allele specific surrogate peptide;
- a station for analyzing the at least one allele specific surrogate peptide by mass spectrometry to detect the at least one allele specific surrogate peptide;
- and,
- a station for determining the genotype of the gene of interest based on results of the of the at least one allele specific surrogate peptide.
The system of claim 20, wherein the station for analyzing the at least one allele specific surrogate peptide by mass spectrometry comprises a tandem mass spectrometer.
Dependencies are inferred from the claim text.