Open AccessTargeted proteomics analysis of plasma proteins using recombinant protein standards for addition only workflows
Author(s) -
David Kotol,
Andreas Hober,
Linnéa Strandberg,
Anne-Sophie Svensson,
Mathias Uhlén,
Fredrik Edfors
Publication year - 2021
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/btn-2021-0047
Subject(s) - proteomics , chaotropic agent , proteome , recombinant dna , mass spectrometry , chemistry , chromatography , computational biology , blood proteins , bioinformatics , biology , biochemistry , gene
Targeted proteomics is an attractive approach for the analysis of blood proteins. Here, we describe a novel analytical platform based on isotope-labeled recombinant protein standards stored in a chaotropic agent and subsequently dried down to allow storage at ambient temperature. This enables a straightforward protocol suitable for robotic workstations. Plasma samples to be analyzed are simply added to the dried pellet followed by enzymatic treatment and mass spectrometry analysis. Here, we show that this approach can be used to precisely (coefficient of variation <10%) determine the absolute concentrations in human plasma of hundred clinically relevant protein targets, spanning four orders of magnitude, using simultaneous analysis of 292 peptides. The use of this next-generation analytical platform for high-throughput clinical proteome profiling is discussed.