panoramaProteomicsKonstantinos KalogeropoulosHomo Sapienshttps://panoramaweb.org/surequant_method.urluadk@dtu.dkTechnical University of DenmarkPanoramaPublicTargeted proteomics methods have been greatly improved and refined over the last decade and are becoming increasingly the method of choice in protein and peptide quantitative assays. Despite the tremendous progress, targeted proteomics assays still suffer from inadequate sensitivity for lower abundant proteins and throughput, especially in complex biological samples. These attributes are essential for establishing targeted proteomics methods at the forefront of clinical use. Here, we report an assay utilizing the SureQuant internal standard-triggered targeted method on a latest generation mass spectrometer coupled with an EvoSep One liquid chromatography platform, which displays high sensitivity and a high throughput of 100 samples per day. We demonstrate the robustness of this method by quantifying proteins spanning six orders of magnitude in human wound fluid exudates, a biological fluid that exhibits sample complexity and composition similar to plasma. Among the targets quantified were low-abundance proteins such at tumor necrosis factor A and interleukin 1-β, highlighting the value of this method in the quantification of trace amounts of invaluable biomarkers that were until recently hardly accessible by targeted proteomics methods. Taken together, this method extends the toolkit of targeted proteomics assays and will help to drive forward mass spectrometry-based proteomics biomarker quantification.High-Throughput and High-Sensitivity Biomarker Monitoring in Body Fluid by Fast LC SureQuant IS-Targeted Quantitation.Kalogeropoulos Konstantinos K, Savickas Simonas S, Haack Aleksander M AM, Larsen Cathrine A CA, Mikosiński Jacek J, Schoof Erwin M EM, Smola Hans H, Bundgaard Louise L, Auf dem Keller Ulrich UfalseHigh-throughput and high-sensitivity biomarker monitoring in body fluid by fast LC SureQuant™ IS targeted quantitationTargeted proteomics methods have been greatly improved and refined over the last decade and are becoming increasingly the method of choice in protein and peptide quantitative assays. Despite the tremendous progress in targeted proteomic assays, they still suffer from inadequate sensitivity for lower abundant proteins and throughput, especially in complex biological samples. These attributes are essential for establishing targeted proteomics methods at the forefront of clinical use. Here, we report an assay utilizing the SureQuantTM internal standard triggered targeted method on a contemporary mass spectrometer coupled with an EvoSep One liquid chromatography platform, which displays markedly enhanced sensitivity and a high throughput of 100 samples per day. We demonstrate the robustness of this method by quantifying proteins ranging six orders of magnitude in human wound fluid exudates, a biological fluid that exhibits sample complexity and composition similar to plasma. Among the targets quantified were low abundant proteins such at TNFA and IL1B, highlighting the value of this method in the quantification of trace amounts of invaluable biomarkers that were until recently outside of the sphere of quantification with targeted proteomics methods. Taken together, this method provides an excellent addition to the toolkit of targeted proteomic assays and has the potential to be a centerpiece in biomarker quantification studies.Tue Oct 07 00:00:00 GMT+01:00 2025PXD032305960639442693