Data-independent acquisition (DIA)
allows comprehensive proteome
coverage, while it also potentially works as a unified protocol to
determine a multitude of proteins found in blood. Because of its high
specificity, mass spectrometry may greatly reduce the interference
observed in other assays to evaluate blood markers. Here, we combined
DIA with volumetric absorptive microsampling (VAMS) and automated
proteomics sample processing in a platform to assess clinical markers.
As a proof of concept, we evaluated two hemoglobin-related biomarkers:
the glycated hemoglobin (HbA1c) and hemoglobin (Hb) variants. HbA1c
by DIA showed good correlation with the reference method, but method
imprecision did not meet the quality requirement for this biomarker.
We developed a strategy to identify Hb variants based on a customized
database combined with a workflow for DIA data extraction and rigorous
peptide evaluation. Data are available via ProteomeXchange with identifier
PXD029918.