Reproducible protein quantitation of 270 human proteins at increased depth using nanoparticle-based fractionation and reaction monitoring mass spectrometry with stable isotope-labelled internal standards
Gaither C, Popp R, Gajadhar AS, Borchers CH. Reproducible protein quantitation of 270 human proteins at increased depth using nanoparticle-based fractionation and multiple reaction monitoring mass spectrometry with stable isotope-labelled internal standards. Analyst. 2025 Jan 13;150(2):353-361. doi: 10.1039/d4an00967c. PMID: 39670628.
- Organism: Homo sapiens
- Instrument: 6495C Triple Quadrupole LC/MS
- SpikeIn:
Yes
- Keywords:
plasma proteomics, LC/MRM-MS, fractionation, absolute quantitation, Seer Proteograph
-
Lab head: Christoph Borchers
Submitter: Claudia Gaither
When using a mix of 274 light synthetic peptide standards as surrogates for 270 human plasma proteins, as well as stable isotope-labelled standards as normalizers for targeted quantitative analysis by LC/MRM-MS, the Seer Proteograph allowed for the enrichment and absolute quantitation of up to an additional 44% of protein targets (median) as well as improved reproducibility compared to a traditional proteomic workflow with no fractionation (median 8.3% vs. 13.1% CV). As the Proteograph technology gains popularity, establishing Proteograph-specific protein reference ranges and comparisons to methods like ELISA and LC/MRM-MS may be explored to enable absolute quantification of plasma proteins based on Proteograph data.
Created on 7/5/24, 3:57 PM