Data-independent acquisition (DIA) has emerged as the preferred analysis method for many proteomics researchers, as it provides an unbiased, global view of the proteome with all peptides within a defined mass-to-charge (m/z) window subject to fragmentation. With the introduction of Zeno SWATH DIA on the ZenoTOF 7600 system, researchers now have access to unparalleled speed, sensitivity and accuracy for the identification and quantitation of proteins, particularly when quantitation is performed on MS/MS fragments.
This poster demonstrates the performance of targeted high-resolution MS/MS (MRMHR) and Zeno SWATH DIA for the quantitation of peptides in a complex matrix using microflow liquid chromatography (LC). A lower limit of quantitation (LLOQ) of 0.02 fmol/µL (0.1 fmol on column) was demonstrated for alcohol dehydrogenase (ADH1) protein digest from yeast spiked into a background of K562 cell lysate tryptic digest. Excellent linearity was observed for both the MRMHR and Zeno SWATH DIA methods, with a linear dynamic range of 3.0 and 2.5 orders of magnitude, respectively. Scheduled MRMHR (sMRMHR) improved the LLOQ by a factor of 3 on average relative to Zeno SWATH DIA. These results highlight the versatility and quantitative capabilities of the ZenoTOF 7600 system, which make it ideal for the identification and robust quantitative verification of protein biomarkers.