期刊
JOURNAL OF PROTEOME RESEARCH
卷 21, 期 6, 页码 1408-1417出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.jproteome.2c00014
关键词
absolute quantification of proteins; MS Western; QconCAT; targeted quantitative proteomics; cerebrospinal fluid; neurodegeneration; neuroinflammation
资金
- Max Planck Society
The FastCAT method is a fast approach for the absolute quantification of clinically relevant proteins, which improves the accuracy, precision, and dynamic range of quantification without the need for purification of chimeric proteins or the use of gel electrophoresis techniques.
Absolute (molar) quantification of clinically relevant proteins determines their reference values in liquid and solid biopsies. The FastCAT (for Fast-track QconCAT) method employs multiple short (<50 kDa), stable-isotope labeled chimeric proteins (CPs) composed of concatenated quantotypic (Q)-peptides representing the quantified proteins. Each CP also comprises scrambled sequences of reference (R)-peptides that relate its abundance to a single protein standard (bovine serum albumin, BSA). FastCAT not only alleviates the need to purify CP or use sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) but also improves the accuracy, precision, and dynamic range of the absolute quantification by grouping Q-peptides according to the expected abundance of the target proteins. We benchmarked FastCAT against the reference method of MS Western and tested it in the direct molar quantification of neurological markers in human cerebrospinal fluid at the low ng/mL level.
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