Absolute (molar) quantification of proteins provides the analytical rationale for system-level modelling of diverse molecular mechanisms. 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 sequences of reference (R-) peptides that relate its abundance to a single protein standard (BSA). FastCAT not only alleviates the need in purifying CP or using 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 target proteins.
Ignacy Rzagalinski, Viditha Rao, Aliona Bogdanova, Lena Hersemann, Andrej Shevchenko
Targeted Absolute Quantification of Protein Biomarkers in Cerebrospinal Fluid by FastCAT.
Methods Mol Biol, 2914 65-74 (2025)DOI: 10.1007/978-1-0716-4462-1_6
Ignacy Rzagalinski, Aliona Bogdanova, Bharath Kumar Raghuraman, Eric R Geertsma, Lena Hersemann, Tjalf Ziemssen, Andrej Shevchenko
FastCAT Accelerates Absolute Quantification of Proteins Using Multiple Short Nonpurified Chimeric Standards.
J Proteome Res, 21(6) 1408-1417 (2022) DOI: 10.1021/acs.jproteome.2c00014
Contact: rzagalinmpi-cbg.de