Selected reaction monitoring assays
Inventors
Kearney, Paul Edward • Li, Xiao-Jun • Hayward, Clive • Geeraerts, Miguel Dominguez
Assignees
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Publication Number
US-10534002-B2
Publication Date
2020-01-14
Expiration Date
Abstract
Provided herein are methods for developing selected reaction monitoring mass spectrometry (LC-SRM-MS) assays.
Core Innovation
The document discloses LC-SRM-MS assay development and analysis workflows for quantifying a multiplex panel of proteins using selected tryptic peptides. The approach is based on selecting peptide precursor/product transitions and using retention time and transition behavior to support assay specificity for multiplex measurement.
Peptide selection is governed by defined criteria, including tryptic peptide constraints, peptide mass ranges, and transition selection rules. Interfering transitions are avoided by applying an offset criterion between interfering transitions and by selecting transitions with appropriate b/y transition ion behavior and intensity.
The disclosed work further supports specificity through transition correlation and selection of unique, non-modified peptides. The document describes compositions of isolated synthetic peptides, including peptides with corresponding SEQ ID numbers and optional isotopically labeled internal standards, within LC-SRM-MS workflows on triple quadrupole LC-SRM-MS platforms.
Claims Coverage
The independent claims cover three inventive features: a fixed six-peptide composition, a panel with at least two isolated synthetic peptides for each of TSP1, ALDOA, FRIL, LG3BP, and C163A, and a panel with at least five isolated synthetic peptides for each of the same proteins.
Specific isolated synthetic peptide set
A composition comprising isolated synthetic peptides said composition comprising ALQASALK (SEQ ID NO: 58), GFLLLASLR (SEQ ID NO: 1644), LGGPEAGLGEYLFER (SEQ ID NO: 596), VEIFYR (SEQ ID NO: 941), INPASLDK (SEQ ID NO: 153) and TSYQVYSK (SEQ ID NO: 155).
At least two isolated synthetic peptides per listed protein
A composition comprising at least 10 isolated synthetic peptides said composition comprising at least 2 isolated synthetic peptides of each TSP1, ALDOA, FRIL, LG3BP, and C163A, selected from the listed peptide set.
At least five isolated synthetic peptides per listed protein
A composition comprising at least 25 isolated synthetic peptides said composition comprising at least five isolated peptides of each TSP1, ALDOA, FRIL, LG3BP, and C163A, selected from the listed peptide set.
Coverage is defined by a fixed composition of six specified isolated synthetic peptides and by larger peptide panels for TSP1, ALDOA, FRIL, LG3BP, and C163A with minimum peptide-count requirements.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Not explicitly described in patent.
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