Optimization of bottom-up proteomic sample preparation by design of experiments for absolute quantification of 257 proteins in human plasma using UPLC-MRM-MS
We successfully reduced protein digestion time from 18 hours (overnight) to 4 hours while maintaining relative digestion efficiency. We improved the sensitivity of the assay via the optimized workflow and were able to quantify proteins that previously fell below the LLOQ. These advancements, combined with automation, provide a practical, efficient, and reproducible workflow suitable for clinical research.
Authors
- Kristrun Yr Holm
- Finnur Freyr Eiríksson
- Margrét Þorsteinsdóttir (ORCID: https://orcid.org/0000-0002-0649-8503)
- Christoph H. Borchers (ORCID: https://orcid.org/0000-0003-2394-6512)
- Yassene Mohammed (ORCID: https://orcid.org/0000-0003-3265-3332)
- Sigríður Klara Böðvarsdóttir (ORCID: https://orcid.org/0000-0001-8799-8018)
- Kari Arnarson
- Valdis Gunnarsdottir Thormar (ORCID: https://orcid.org/0009-0005-2874-4081)
Institutions
- University of Iceland (IS)
- Leiden University Medical Center (NL)
- McGill University (CA)
Publication Details
- Journal
- Expert Review of Proteomics
- Published
- 2025-05-04
- DOI
- https://doi.org/10.1080/14789450.2025.2504994
- Citations
- 1
- Primary Topic
- Advanced Proteomics Techniques and Applications
- Type
- article
- Field-Weighted Citation Impact
- 0.76