The 60 samples per day method
More than 4000 proteins in 21 minutes
The 60 SPD method is one of our most popular standard methods as it provides an excellent balance between throughput and depth.
The method is ideal for deep fractionated proteomes as well as single-shot analyses, where more than 4000 proteins are routinely identified with DIA.
ENHANCED PROTEOME COVERAGE WITH THE PERFORMANCE COLUMN
The performance column is for exceptional restult and available for the 60 samples per day method. It is packed with 1.5 μM C18 beads, which puts some demands on usage such as requiring elevated column temperature control to manage system backpressure.
The smaller beads reduces peaks capacity of 60%. Consequently, the proteome coverage is enhanced, and the column provides a robust and competetive commercial option – on the same level as the home-pulled and packed options, but with better reproducibility and life-time.
The boost in sensitivity is particularly noticeable for lower loads, where peptide and protein coverage is improved with 100%.
QUANTITATIVE RAT ORGAN PROTEIN ATLAS BY TMT AND dia-lfq
In this publication, the Olsen Group from the Novo Nordisk Foundation Center for Protein Research, Denmark, perform a comprehensive rat tissue specific protein atlas. They compare tandem mass tags (TMT11-plex) with DIA-based label-free quantification of 12 tissues covering the major mammalian organs from three individual male rats.
The spectral library generated, represents 213,000 unique peptides covering 12,909 protein-coding genes and should represent a resource for the proteomics community.
Unsupervised hierarchical clustering of relative protein abundances across tissues reveal very similar protein expression patterns between DIA-FAIMS and TMT experiments, suggesting that both quantification strategies provides very similar biological insights.
UNLEASHING THE TRUE POWER OF DIA ACQUISITION WITH SHORT GRADIENTS
In this case study, Ben C. Collins from Queen’s University, Belfast and Ting Huang, Northeastern University, Boston discuss the importance of fast and standardized workflows
Analysis speed can be increased by one order of magnitude with the Evosep One, reducing analysis time from roughly 10 days to a little more than 1 day without significant loss of information at the protein complex level. This enables novel applications for profiling proteome organization dynamics.
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FAST AND REPRODUCIBLE PHOSPHOPROTEOMICS
This application note describes an automated workflow for protein digestion as well as enrichment of phosphopeptides using MagReSyn magnetic beads for the entire workflow.
By employing a streamlined and automated workflow for phosphopeptide enrichment in combination with fast and robust chromatographic performance delivered by the Evosep One, we offer a workflow for routine use in large clinical studies, which can also be easily extended to tissue samples.
The use of automation in the entire workflow eliminates the hands-on challenges and ensures excellent reproducibility when handling a large number of samples.
The five standard methods
With the five standard methods, the Evosep One covers a range of use cases from comprehensive proteome analysis with fractionation strategies to ultra high-throughput single-shot analysis. You device how fast you want to go.
The highest peak capacity per minute with our fastest method for ultra-high throughput analysis.
Excellent sequencing speed with most peptides per minute among our standard methods.
A great “in-between” method with run-to-run reproducibility of just one second.
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EVOSEP ONE – A SEPARATION TOOL DESIGNED FOR CLINICAL OMICS
IMPROVE AND ENHANCE YOUR PROTEOME COVERAGE WITH THE PERFORMANCE COLUMN
Do you want to know how our users are already taking advantage of the 300 Samples per day method? Find all publications from users and collaboration partners here. Visit our Literature Room for a full overview.
|Title||Subject||Material||Year||Summary||Institute||Evosep method||MS instrumentation||Learn More|
|Deciphering the LRRK code: LRRK1 and LRRK2 phosphorylate distinct Rab proteins and are regulated by diverse mechanisms||Parkinson's disease, Targeted workflow||Publication||2020||This publication from University of Dundee led by Dario R Alessi, describes how LRRK1, a less studied homologue of LRRK2 regulates growth factor receptor trafficking and osteoclast biology reinforcing that the LRRK enzymes have evolved as major regulators of Rab biology in Parkinson’s disease.||University of Dundee, United Kingdom. Johann Wolfgang Goethe University, Germany. Michael J Fox Foundation for Parkinson’s Research, United States.||60 SPD||Thermo Q Exactive HF-X|
|Development of a multiplexed targeted mass spectrometry assay for LRRK2-phosphorylated Rabs and Ser910/Ser935 biomarker sites||Clinical research, Parkinson's disease, Targeted workflow||Publication||2020||This publication from University of Dundee led by Dario R Alessi, describes the validation and development of a new, multiplexed targeted assay that enables the relative quantification of the key components of the LRRK2 pathway, defining the impact of LRRK2 inhibitors and Parkinson’s disease causing mutations.||University of Dundee, United States. Medical University of Vienna, Austria.||30 SPD, 60 SPD||Thermo Orbitrap Exploris 480|
|diaPASEF: parallel accumulation–serial fragmentation combined with data-independent acquisition||diaPASEF, Technology||Publication||2020||A collaboration led by the Mann group at the Max Planck Institute for Biochemistry, Münich describes the development of the latest addition to the PASEF technology. The diaPASEF workflow combines the advantages of DIA such as high degree of reproducibility and data completeness with the effective PASEF technology.||Max Planck Institute of Biochemistry, Martinsried, Germany. Bruker Daltonik, Germany. ETH Zürich, Switzerland. University of Belfast, United Kingdom. University of Toronto, Canada.||100 SPD, 200 SPD, 60 SPD||Bruker timsTOF Pro|
|High throughput proteome and phosphorpoteome sample processing coupled to fast gradient DIA||DDA, DIA, FAIMS, Phosphorylation, PTM, Technology, Tissue, TMT||Publication||2020||High throughput workflows for proteomics||Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Denmark||100 SPD, 200 SPD, 60 SPD||Thermo Orbitrap Exploris 480|
|Peptidomic analysis of urine from youths with early type 1 diabetes reveals novel bioactivity of uromodulin peptides in vitro||Clinical research, Peptidomics, Targeted workflow, Urine||Publication||2019||Urinary peptidomics in early type 1 diabetes||SickKids Proteomics, Analytics, Robotics, and Chemical Biology Centre, Canada||60 SPD||Thermo Q Exactive HF-X|
|New Orbitrap Exploris 480 Mass Spectrometer Coupled with Evosep One||DDA, DIA, FAIMS, Technology||Brochure||2020||Brochure of Exploris 480 with Evosep data from piece no 1||Thermo Fisher Scientific, United States||100 SPD, 200 SPD, 60 SPD||Thermo Orbitrap Exploris 480|
|Development of a robust and reprodusible method for detection of citrullination in complex samples||Bacteria, Citrullination, DDA, PTM||Poster||2018||Detection of citrullination in complex samples||University of Southern Denmark, Denmark||60 SPD||Thermo Orbitrap Fusion Lumos|
|How to get Speed and Depth in yout Host Cell Protein||Biopharma, DDA, HCP||Video||2018||Bruker Daltonics, Germany||60 SPD||Bruker timsTOF Pro|
|A Novel LC System Embeds Analytes in Pre-formed Gradients for Rapid, Ultra-Robust Proteomics||DDA, DIA, Plasma, Technology||Publication||2018||Description of Evosep||Max Planck Institute of Biochemistry, Martinsried Germany and Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Denmark||60 SPD||Thermo Q Exactive HF-X|
|Robust and Reprodusible Protein Quantification in Plasma Using the Evosep One and the Agilent 6495 Triple Quadrupole LC/MS||Plasma, Targeted workflow, Technology||Application note||2020||Agilent feasibility study||Agilent Technologies, United States||60 SPD||Agilent 6495 Triple Quadrupole|
|Development of a Novel LC Concept for Clinical Proteomics||DDA, Plasma, Technology||Poster||2018||Description of Evosep One||Max Planck Institute of Biochemistry, Martinsried and Bruker Daltonics, Germany||60 SPD||Thermo|
|Increasing proteome coverage using cysteine-specific DIA Mass spectrometry – Cys-DIA||DIA, PTM||Publication||2020||DIA on cysteine-specific peptides||University of Southern Denmark||60 SPD||Thermo Q Exactive HF-X|
|Evosep One Enables Robust Deep Proteome Coverage Using Tandem Mass Tags While Significantly Reducing Instrument Time||DDA, Lung, Tissue, TMT||Video||2019||Comparison study with TMT, same coverage in shorter time||SickKids Proteomics, Analytics, Robotics, and Chemical Biology Centre, Canada||30 SPD, 60 SPD||Thermo Orbitrap Fusion Lumos|
|A network of RNA-binding proteins controls translation efficiency to activate anaerobic metabolism||DDA, TMT||Publication||2020||SickKids Proteomics, Analytics, Robotics, and Chemical Biology Centre, Canada||60 SPD||Thermo Orbitrap Fusion Lumos|
|Improving proteome coverage and peptide identification rates in short LC gradients||DDA, DIA, FAIMS, Phosphorylation, PTM, Technology||Video||2020||Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Denmark||100 SPD, 200 SPD, 60 SPD||Thermo Orbitrap Exploris 480|
|High-throughput proteomics with Evosep One||DDA, DIA, FAIMS, Phosphoproteomics, PTM, Technology||Video||2020||Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Denmark||100 SPD, 200 SPD, 60 SPD||Thermo Orbitrap Exploris 480|
|Rapid proteome analyses using the Evosep One||DDA, DIA, Phosphorylation, PTM||Video||2018||Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Denmark||60 SPD||Thermo Q Exactive HF|
|Sensitive, Rapid, Robust and Reproducible Workflow for Host Cell Protein Profiling in Biopharmaceutical Process Development||Biopharma, DDA, HCP||Publication||2020||HCP analysis in Biopharma||Glaxo Smith Kline, Unites States||30 SPD, 60 SPD||Thermo Orbitrap Fusion Lumos|
|Using Artificial Intelligence on Ultrafast LC-MSMS-DIA runs for Bacterial Identification in Urine||Bacteria, Clinical research, DIA||Video||2019||View the recording of our HUPO 2019 lunch seminar, where Florence Roux-Dalvai from Québec Research Center, Canada presents a new strategy for bacterial species identification in urinary tract infection using artificial intelligence on ultrafast LCMS DIA runs.||CHU Québec Université Laval Research Center, Canada||100 SPD, 200 SPD, 60 SPD||Thermo Orbitrap Exploris 480|
|High-throughput 4D-Proteomics – Application of dia-PASEF® and the Evosep One for short gradients||DIA, Technology||Application note||2020||diaPASEF app note||Bruker Daltonics, Germany||100 SPD, 200 SPD, 300 SPD, 60 SPD||Bruker timsTOF Pro|
|Fast and reproducible phosphoproteomics using MagReSyn® Amine and Tim-IMAC HP magnetic beads and the Evosep One||Phosphorylation, PTM, Technology||Application note||2020||Read this application note to learn about the automated workflows for sample preparation developed by the Olsen group in collaboration with Resyn Biosciences. Protein digestion and phosphoenrichment is fully automated for up to 96 samples in parallel with the Kingfisher Flex robot from Thermo Scientific.||Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Denmark||60 SPD||Thermo Orbitrap Exploris 480|
|Consistency, consistency – Automated Sample Prep for Translational Proteomics||Clinical research, Plasma||Video||2020||In this webinar, Emily Chen, Sr. Director at the Thermo Fisher Precision Medicine Science Center presents an automated, robust and scalable sample preparation pipeline for large-scale clinical research samples.||Thermo Fisher Scientific, United States||60 SPD||Thermo|
|An Ultra High-Throughput Plasma Protein Profiling (uHTPPP) Workflow Using a Modified Quadrupole-Orbitrap Mass Spectrometer||Clinical research, DDA, Lung, Plasma, Serum, Tissue||Application note||2020||In this application note, the group of Emily Chen at the Thermo Fisher Precision Medicine Science Center describes a high-throughput plasma and serum proteomics analysis workflow for large population cohort studies that utilizes a standardized sample preparation method, high-throughput data acquisition, and easy to implement QC standard.||Thermo Fisher Scientific, United States||30 SPD, 60 SPD||Thermo Orbitrap Exploris 240|
|Scalable and Automated Plasma Workflow Based on the Thermo Scientific Q Exactive HF-X MS Platform||Clinical research, DDA, Lung, Plasma, Serum||Application note||2019||In this application note, the group of Emily Chen at PMSC describes a high-throughput plasma and serum proteomics analysis workflow for large population cohort studies that utilizes a standardized sample preparation method, high-throughput data acquisition, and easy to implement QC standard.||Thermo Fisher Scientific, United States||100 SPD, 30 SPD, 60 SPD||Thermo Q Exactive HF-X|
|Advancing Interactomics Workflows with 4D-Proteomics on the timsTOF Pro||DDA, Yeast||Video||2020||In this webinar organized by Bruker, André Michaelis from Matthias Mann’s group presents his large-scale yeast interactome study. He describes a robust platform for affinity purification and an LCMS workflow with Evosep One and timsTOF Pro with more than 8000 LCMS runs completed in just 20 weeks.||Max Planck Institute of Biochemistry, Martinsried, Germany||60 SPD||Bruker timsTOF Pro|
|Oncogenic Mutations Rewire Signaling Pathways by Switching Protein Recruitment to Phosphotyrosine Sites||DIA, Liver, Lung, Pulldown, Tissue||Publication||2019||In this publication, the Olsen Group have combined DIA with short LC gradients to describe how cancer mutations close to tyrosine phosphorylation sites in the EGF receptor rewire signaling pathways by switching protein interactors. The performed 1200 pulldown experiments in just 20 days of LC-MS instrument time.||Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Denmark||60 SPD||Thermo Q Exactive HF-X|
|timsTOF Pro with PASEF and Evosep One: Maximizing throughput, robustness and analytical depth for shotgun proteomics||DDA, Technology||Application note||2018||Read this application note to learn more about the timsTOF Pro with PASEF and the Evosep One for maximizing throughput, robustness and analytical depth for shotgun proteomics. Increased peak capacity via trapped ion mobility spectrometry allows the analysis of 200 samples/day, at formidable analytical depth, with unparalleled robustness.||Bruker Daltonics, Germany||100 SPD, 200 SPD, 60 SPD||Bruker timsTOF Pro|
|Multi-level proteomics reveals host-perturbation strategies of SARS-CoV-2 and SARS-CoV||Clinical research, Covid-19, DIA, PTM||Publication||2020||This publication describes the molecular functions of viral proteins and their interactions with the host proteome of SARS-CoV-2. The impact of viral infection on the proteome and phosphoproteome were analyzed in a time-resolved manner by DIA. The analysis revealed key pathways perturbed during the infection identifying potential vulnerable points of SARS-CoV-2.||Max Planck Institute of Biochemistry, Martinsried, Germany||30 SPD, 60 SPD||Thermo Q Exactive HF-X|
|Evosep One Enables Robust Deep Proteome Coverage Using Tandem Mass Tags While Significantly Reducing Instrument Time||Offline fractionation, Tissue, TMT||Video||2019||Comparison study with TMT, same coverage in shorter time||SickKids Proteomics, Analytics, Robotics, and Chemical Biology Centre, Canada||30 SPD, 60 SPD||Thermo Orbitrap Fusion Lumos|
|Increasing Throughput: From Pre-Clinical Models To Protein Complexes||DIA, Large-scale||Video||2019||Comparison study with >400 samples, same coverage in shorter time||ETH Zürich, Switzerland||60 SPD||Sciex|
|Fast and Robust Proteome Screening Platform Identifies Neutrophil Extracellular Trap Formation in the Lung in Response to Cobalt Ferrite Nanoparticles||DDA, Lung, Tissue||Publication||2020||In vivo study of biocompatibility of magnetic nanoparticles. Toxicity of magnetic metal oxide nanoparticles on the respiratory system was examined in vivo by single intratracheal instillation in mice. Bronchoalveolar lavage fluid (BALF) samples were collected for proteome analyses by LC−MS/MS.||University of Southern Denmark||60 SPD||Thermo Orbitrap Fusion Lumos|
|A Compact Quadrupole-Orbitrap Mass Spectrometer With Faims Interface Improves Proteome Coverage In Short Lc Gradients||Automation, DDA, DIA, FAIMS, Offline fractionation, Phosphoproteomics, PTM, Technology, Tissue, TMT||Publication||2020||Test of Orbitrap Exploris with DIA and TMT, proteome and phospho.||Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Denmark||100 SPD, 200 SPD, 60 SPD||Thermo Orbitrap Exploris 480|
|Advanced Host Cell Protein (Hcp) Analysis With Tims Qtof Ms Powers Biopharmaceutical Development||Antibodies, DDA, HCP||Publication||2019||Descriptive piece of HCP analysis on the timsTOF||Bruker Daltonics, Germany||60 SPD||Bruker timsTOF Pro|
|High-Throughput Proteomics Quantification Enabled by Fast LC Separation and Advanced PRM Acquisition||Phosphorylation, PTM, Targeted workflow||Poster||2018||High throughput LC-PRM to monitor the main protein components of AKT/mTOR signaling pathway.||Thermo Fisher Scientific, United States||100 SPD, 200 SPD, 60 SPD||Thermo Q Exactive HF-X|
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