However, the dynamic range of protein concentration in these samples poses a challenge for in-depth proteome quantification. PASEF® is a powerful technology for rapid in-depth and sensitive proteome quantification that is also applicable to biofluids like plasma and serum. We combined PASEF with high-throughput gradient settings for the analysis of hundreds of plasma and serum samples and quantified several hundred protein groups routinely in 21- and 47-minute runtimes. The timsTOF Pro coupled with the Evosep One nanoflow UPLC delivers quantitation with median CV less than 20% and quantified more than 70 known biomarkers in a study investigating over 700 COVID-19 serum samples.