Targeted workflows

Targeted workflows are efficiently used to detect specific peptide ions to verify and validate specific biological hypotheses. 

Combined with recent years technological advancements, this has led to the expansion of targeted strategies, which offer high sensitivity, quantitative accuracy and reproducibility.

Why targeted workflows? 

Targeted proteomics is ideal for hypothesis-based studies of a subset of proteins with high sensitivity, quantitative accuracy and reproducibility.

These workflows have gained in popularity, especially to detect established biomarkers for several applications. An increased focus, along with technological advancements is continuously pushing the limits for high-throughput targeted analysis.

The Evosep One is specifically designed for clinical applications, where robustness and reproducibility are key priorities. With very high retention time reproducibility, it is the ideal front-end solution for robust and high-throughput targeted workflows.

Robust and Reproducible Protein Quantification in Plasma 

In collaboration with Agilent technologies, we highlight the reproducibility, robustness, and analytical sensitivity of MRM-based LC/MS analysis of human plasma proteins using the Agilent 6495 triple quadrupole LC/MS with the Evosep One.

The results demonstrate superior retention time reproducibility over a 12-day analysis period, and equivalent sensitivity before and after robustness testing, underlining the suitability of the system for high-throughput protein quantification.

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A PERSONALIZED MS-BASED ASSAY TO MONITOR MULTIPLE 2 MYELOMA DISEASE 

In collaboration with Rapid Novor and University of Waterloo, the Trudel group describes the development of a two step non-invasive MS-based assay, called EasyM.

This assy  is used to assess minimal residual disease, a measure of depth of remission to treatment, which has become an important parameter in assessing the disease burden in multiple myeloma.

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high-throughput doping control with the Evosep One using the High organic method

Doping is a significant problem in horse racing and sport testing laboratories run tens of thousands of samples per year. Anabolic androgenic steroids (AAS) represent one of the most frequent detected classes of prohibited substances in doping controls due to their long-lasting beneficial effects on performance.

In collaboration with LCH-GIE France, we have developed the High Organic Method, specifically designed to detect stanozolol and its phase I metabolites in challenging matrices such as horse urine and hair. This is the first Evosep One method for small-molecule analysis.

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Targeted workflows have gained in popularity, especially to detect established biomarkers for several applications. An increased focus, along with technological advancements is continuously pushing the limits for high-throughput targeted analysis.

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More research with targeted workflows

Here you can see publications available on targeted workflows featuring Evosep One. For a full overview of publications published using the Evosep One Technology visit our Literature room here

TitleSubjectMaterialYearSummaryInstituteEvosep methodMS instrumentationLearn More
A micro-flow, high-pH, reversed-phase peptide fractionation and collection system for targeted and in-depth proteomics of low-abundance proteins in limiting samples, 2023

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Proteogenomic analysis of chemo-refractory high- grade serous ovarian cancer, , , 2023

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Development of a Standardized Microflow LC Gradient to Enable Sensitive and Long-Term Detection of Synthetic Anabolic-Androgenic Steroids for High-Throughput Doping Controls, 2021This publication led by the Bailly-Chouriberry group at GIE-LCH, Laboratoire Des Courses Hippiques in France presents the expanded detection window of stanozolol in complex matrices such as equine urine and hair using the High Organic method for more hydrophobic compounds.

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Targeted Workflows with Evosep One, , 2021During this webinar, Michael MacCoss (University of Washington) and Vivian Delcourt (GIE-LCH) share how they use the Evosep One for targeted workflows.

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A personalized mass spectrometry-based assay to monitor M-protein in multiple myeloma patients (EasyM), , , 2021This publication by the Trudel group describes the development of a non-invasive MS-based assay, called EasyM, which is used to assess minimal residual disease, a measure of depth of response to treatment, which has become an important parameter in assessing the disease burden in multiple myeloma., ,

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Mass Spectrometry Provides a Highly Sensitive Noninvasive Means of Sequencing and Tracking M-Protein in the Blood of Multiple Myeloma Patients, , , 2021The amino acid sequence of the M-protein for multiple myeloma is unique compared to the polyclonal antibodies in patients’ blood. This publication from Rapid Novor describes a targeted MS/MS assay to detect and quantify the unique M-protein sequence in serum samples from multiple myeloma patients.

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Rapid and Accurate Detection of Aminoglycoside-Modifying Enzymes and 16S rRNA Methyltransferases by Targeted Liquid Chromatography-Tandem Mass, 2021This publication by the Dekker group describes the development of an accurate and rapid targeted LC-MS/MS assay based on parallel reaction monitoring for detection of the most prevalent aminoglycoside-modifying enzymes and 16S rRNA methyltransferases in Escherichia coli and Klebsiella pneumoniae that confer resistance to aminoglycosides.

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R1441G but not G201S Mutation Enhances LRRK2 Mediated Rab10 phosporylation in human Peripheral blood neutrophils, , 2021This publication from University of Dundee led by Esther Sammler, describes that in vivo LRRK2 dependent

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Deciphering the LRRK code: LRRK1 and LRRK2 phosphorylate distinct Rab proteins and are regulated by diverse mechanisms, 2020This 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.

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Development of a multiplexed targeted mass spectrometry assay for LRRK2-phosphorylated Rabs and Ser910/Ser935 biomarker sites, , 2020This 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.,

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Peptidomic analysis of urine from youths with early type 1 diabetes reveals novel bioactivity of uromodulin peptides in vitro, , , 2019Urinary peptidomics in early type 1 diabetes

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Robust and Reprodusible Protein Quantification in Plasma Using the Evosep One and the Agilent 6495 Triple Quadrupole LC/MS, , 2020Agilent feasibility study

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Targeted assay development for quantifying protein half-life over a range of mouse tissues, , 2019Protein turnover

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Development of mass spectrometry-based targeted assay for direct detection of novel SARS-CoV-2 coronavirus from clinical specimens, , , 2020This publication by the Akhilesh Pandey group describes a targeted FAIMS-PRM assay on an Orbitrap Exploris 480 MS. More than 700 nasopharyngeal swab samples were analyzed with high specificity and comparable sensitivity to the gold standard RT-PCR method for the diagnosis of SARS-CoV-2.,

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Accurate Detection Of The Four Most Prevalent Carbapenemases In E. Coli And K. Pneumoniae By High-Resolution Mass Spectrometry, 2019“Proof of concept” study showing the potential of targeted high resolution MS in detecting different resistance mechanisms. Accurate detection of KPC, OXA-48-like, NDM and VIM carbapenemases.

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High-Throughput Proteomics Quantification Enabled by Fast LC Separation and Advanced PRM Acquisition, , 2018High throughput LC-PRM to monitor the main protein components of AKT/mTOR signaling pathway., ,

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Fast Confirmatory Analysis of Rhuepos in Plasma for Horseracing Doping Control, , 2019This application note describes a rapid targeted analysis monitoring two tryptic rHuEPO peptides in plasma for horseracing doping control. The results illustrate the promising capabilities of the Evosep One innovative technology for the analysis of protein/peptide-based drugs in the context of drug testing.

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Evosep One solves the proteomics dilemma – covering high throughput analysis and proteome depth
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end-to-end, fully automated digestion protocol and evotip pure workflow on the opentrons ot-2
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