主权项 |
1. A method for detecting the presence of one or more fusion proteins within a biological sample, comprising:
a) providing:
a biological sample, wherein said biological sample is a blood sample and/or a tissue sample,one or more distinct groups of labeled peptides, wherein each distinct group of labeled peptides is specific for a particular fusion protein to be detected, wherein the labeled peptides within each distinct group have labeled peptides having an amino acid sequence that spans the fusion junction of said particular fusion protein encoded by a fusion of a first nucleic acid to a second nucleic acid,
wherein said labeled peptides within each distinct group comprise peptides having one isotopically labeled amino acid residue and peptides having two isotopically labeled amino acid residues,wherein said amino acid sequence of said labeled peptides within each distinct group is identical with an amino acid sequence of a particular fusion protein peptide fragment generated through digestion of said particular fusion protein with a protease,one or more proteases known to generate upon digestion of said two or more fusion proteins to be detected particular fusion protein peptide fragments, b) digesting said biological sample with said one or more proteases resulting in generation of biological sample-based peptide fragments, c) combining said one or more distinct groups of labeled peptides with said generated biological sample-based peptide fragments, d) purifying said labeled peptides combined with said generated biological sample-based peptide fragments, e) conducting a multiple reaction monitoring mass-spectrometry (MRM-MS) analysis of said purified biological sample-based peptide fragments combined with said labeled peptides, and f) analyzing the results of said MRM-MS analysis, wherein said analyzing comprises detecting the presence or absence of a biological sample-based fusion protein peptide fragment having an amino acid sequence identical with the amino acid sequence for one of the distinct groups of labeled peptides specific for a particular fusion protein, wherein detected co-elution of
i) a biological sample-based fusion protein peptide fragment having an amino acid sequence identical with the amino acid sequence for one of the distinct groups of labeled peptides specific for a particular fusion protein, andii) said amino acid sequence for one of the distinct groups of labeled peptides specific for a particular fusion protein, indicates the presence of said particular fusion protein within said biological sample. |