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Literature room

Application notes, method notes and case studies across the Evosep platform — each summarised with the conditions it was run under and the numbers it reports, so you can find the right document before you download it.

46 documents 7 categories Free to download
Evokit standardized sample preparation kit
Evokit™
Evosep Lupo sample preparation system
Evosep Lupo™
Evotip Pure disposable trap column
Evotip Pure™
Evosep Eno LC separation platform
Evosep Eno™

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Filter by category, or search across titles, summaries and findings — try “carryover”, “plasma”, “single cell”, “500 SPD” or “Astral”.

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Showing all 46 documents

End-to-End Solutions

Automated workflows from sample to Evotip Pure, across liquid handlers and sample types. 8 documents

Application note · AN-020D

A complete, automated Opentrons OT-2 loading protocol for simplified sample loading of Evotip Pure

An Opentrons OT-2 protocol that automates loading of peptide samples onto Evotip Pure, covering runs of 8 to 288 tips. Performance was assessed across peptide input amounts, across every tip position in a full run, and between two separate OT-2 units.

  • ~3,100 proteins identified from 1 ng peptide input, and ~7,000 proteins from 50 ng
  • An average of 2,500 protein groups across all 288 Evotips loaded in a single run
  • Robot-to-robot difference below 10% in identified precursors, averaging ~33,000 precursors
  • Scales from 8 to 288 Evotips per run
Evotip PureAutomationOT-2timsTOF Pro 2Whisper 40 SPD
PDF · 4 pagesDownload →
Application note · AN-027B

Modular sample preparation strategy — fully automated, end-to-end digestion workflow with Evotip Pure loading on the Opentrons OT-2

A hands-off magnetic bead (protein aggregation capture) digestion workflow on the Opentrons OT-2 that ends with samples loaded directly onto Evotip Pure. Throughput, digestion efficiency, reproducibility and input range were characterised on HeLa lysate.

  • 192 samples processed in 6.5 hours, and 384 samples in a single working day per OT-2
  • Over 30,000 precursors identified at 73% digestion efficiency
  • Median CVs of 22% at precursor level and 13% at protein group level
  • 7,000 proteins at 125 ng input, and over 1,000 proteins at 1 ng input
  • A 4-hour room temperature digestion gave results comparable to overnight digestion
Evotip PureAutomationOT-2Sample preptimsTOF Pro 2
PDF · 4 pagesDownload →
Application note · AN-028C

Fully automated, rapid, robust sample loading on Evotip Pure with the Agilent AssayMAP Bravo

An automated Evotip Pure loading protocol on the 96-probe Agilent AssayMAP Bravo. Loading speed, plate position consistency and sensitivity across peptide loads were compared against manual loading.

  • 96 Evotips loaded in approximately 4 minutes
  • Median CV of 8.2% for the automated protocol, against 10% for the manual protocol
  • Nearly 63,000 precursors from a 25 ng load, and close to 3,000 precursors from 100 pg
  • 6,400 proteins identified at the highest load; median CV of 6.5% across plate rows and columns
Evotip PureAutomationAssayMAP BravoZenoTOF 7600timsTOF Pro 2
PDF · 4 pagesDownload →
Application note · AN-030C

Fully automated, end-to-end workflow for neat and deep plasma profiling

Neat plasma profiling compared with MagNet-enriched deep plasma profiling in an automated OT-2 workflow feeding Evotip Pure and Evosep One. Depth, throughput, precision and coverage of FDA-approved biomarkers were assessed on an Orbitrap Astral.

  • Close to 1,500 protein groups from neat plasma, rising to more than 5,000 with the MagNet workflow
  • Up to 192 samples processed in under 10 hours; 100 samples analysed per day
  • Median protein-level CV below 10%
  • 52% (59 of 113) of FDA-approved plasma biomarkers quantified, 72% of those with CV below 20%
PlasmaEvotip PureOT-2MagNetOrbitrap Astral
PDF · 4 pagesDownload →
Application note · AN-031A

Fully automated workflow from raw plasma to ready-to-analyze Evotips

A miniaturized end-to-end OT-2 workflow taking 1 µl of raw plasma through digestion to Evotip Pure without manual intervention. Throughput, digestion completeness, precision and biomarker quantification were evaluated on a timsTOF HT.

  • 192 samples processed in eight hours, from 1 µl plasma input per sample
  • 4,000 precursors corresponding to 400 protein groups, with ~65% fully cleaved peptides
  • Median CV of 13.4%; ~300 proteins per sample, 250+ identified in more than 95% of samples
  • The majority of quantified FDA-approved biomarkers had CV below 20%
PlasmaEvotip PureOT-2MiniaturizationtimsTOF HT
PDF · 4 pagesDownload →
Application note · AN-033B

A sustainable and cost-efficient strategy leveraged by the Evotip Pure — robust digestion workflow for up to 96 samples at a time

An automated protein aggregation capture digestion workflow on the Agilent AssayMAP Bravo, feeding Evotip Pure directly. Digestion duration, reproducibility across a large replicate series and performance across a wide input range were characterised on HeLa lysate.

  • Up to 500 samples processed per day, depending on digestion duration
  • 89% digestion efficiency with overnight digestion, and 65% with a 30-minute digestion
  • 7% CV across 480 technical replicates
  • Over 8,000 proteins identified from 1 µg input; protein CVs below 20% at 10 ng and around 6–7% at 1,000 ng
Evotip PureAutomationAssayMAP BravoSample preptimsTOF HT
PDF · 4 pagesDownload →
Application note · AN-034C

Scalability for high-throughput proteomics — Evotip Pure integration with the Biomek i5 liquid handler

Evotip Pure loading and bead-based sample preparation integrated on a Biomek i5 with a BioShake module, scaling to six plates per run. Cell lysate, neat plasma and MagNet-enriched plasma workflows were each characterised on a timsTOF HT.

  • Up to 576 samples processed in a single run
  • Digestion takes 3 hours for six plates, or 18 hours if run overnight
  • 5,500 proteins per sample from 1 µg input, and 4,900 from 500 ng
  • 500 proteins from neat plasma and 2,700 with Mag-Net enrichment, both at 200 SPD
  • Median protein-level CV below 20% for all workflows
Evotip PureAutomationBiomek i5High-throughputtimsTOF HT
PDF · 4 pagesDownload →
Application note · AN-041A

Evotip Pure ensures ultra-low carryover across 1,100+ plasma samples

The robustness of an automated, miniaturized plasma proteomics pipeline tested over a large clinical cohort on a single platform. Carryover, retention time stability, pressure stability and protein-level precision were tracked across the full series.

  • 1,122 patient plasma samples — 1,392 including controls — analysed in under seven days
  • Carryover below 0.2% across more than 1,100 plasma samples
  • Median protein CV of 15.1% for system suitability controls and 15.6% for plasma proteins
  • Retention time variation below 2% CV across all 88 system suitability controls, and pressure CV below 1% across 1,392 injections
PlasmaEvotip PureOT-2Carryover200 SPDtimsTOF HT
PDF · 4 pagesDownload →

Evosep Eno

Performance, reproducibility and scalability of the Evosep Eno separation platform. 6 documents

Application note · AN-043B

Reproducible, inter-laboratory, scalable and routine LC-MS based proteomics with the Evosep Eno

A three-laboratory study of the complete workflow, each site running its own liquid handler, Evosep Eno and Orbitrap Astral. The design captures the variability that arises from real differences in sample preparation between labs.

  • 5,795 protein groups consistently identified across all three laboratories
  • Approximately 75% quantified with CV below 20%, despite independently prepared lysates
  • Evotip blanks showed less than 0.05% carryover; plate blanks free from cross-well contamination
  • 300 SPD method with 4-hour ambient PAC digestion
Evosep EnoMulti-labReproducibility300 SPDOrbitrap Astral
PDF · 4 pagesDownload →
Application note · AN-044F

Evosep Eno — high sensitivity and quantitative accuracy for plasma proteome analysis at scale

A fully automated plasma workflow coupled to a scheduled MRM assay on the 200 SPD method, monitoring 14 peptides across 11 plasma proteins. The translational case, where accuracy matters more than depth.

  • Detection down to 2 amol, with a linear response spanning 4–5 orders of magnitude
  • 93–110% accuracy (measured versus expected ratio) across a matrix-matched dilution series, R² > 0.99
  • Retention time SD below 0.5 s across 200 consecutive plasma samples
  • Mean peak symmetry 1.1 and FWHM 2.4 s, enabling tight scheduling
Evosep EnoTargeted / MRMPlasma200 SPDXevo TQ-Absolute
PDF · 4 pagesDownload →
Application note · AN-045C

Evosep Eno — new standards for routine, scalable and high-performance LC-MS based proteomics

The six standard Evosep Eno methods characterised end to end on an Orbitrap Astral in DIA mode: how proteome coverage trades against throughput, peak performance across the range, and reproducibility within and between instruments.

  • 9,240 proteins at 30 SPD, 9,028 at 60 SPD, 8,504 at 100 SPD, 7,447 at 200 SPD, 6,663 at 300 SPD and 6,133 at 500 SPD
  • Protein CV of 3.6% at 30 SPD rising to 14.0% at 500 SPD — below 15% across the whole range
  • More than 2,000 protein identifications per minute with the high-performance methods
  • Retention time SD below 0.5 s intra-instrument, ~2 s across eight instruments
Evosep EnoStandard methodsOrbitrap AstralDIAHeLa
PDF · 4 pagesDownload →
Application note · AN-046A

Biological insights at scale with Evosep Eno — 500 samples per day throughput

Eleven sample plates prepared in parallel on a Biomek i7 and analysed with the 500 SPD method, testing whether an automated pipeline holds together across more than a thousand consecutive injections.

  • 1,056 samples from protein lysate to LC-MS result in about 2.5 days
  • Retention time SD of 0.75 s and high-pressure pump stability of 2.5 bar SD across all runs
  • 4,600 protein groups from 200 ng input and 5,500 from 1 µg; inter-plate CVs of 16–18%
  • Consistent 75% digestion efficiency across all eleven plates
Evosep Eno500 SPDAutomationScalabilityBiomek i7
PDF · 4 pagesDownload →
Application note · AN-057A

Whisper™ Zoom drives scalable workflows for high-sensitivity applications

The four Whisper Zoom methods evaluated from 0.25 ng to 200 ng of tryptic HeLa digest, addressing how much throughput must be given up to protect sensitivity at single-cell-equivalent inputs.

  • Nearly 7,000 proteins from just 5 ng of HeLa digest with Whisper Zoom 120 SPD
  • Moving 20 SPD → 120 SPD increases throughput 500% with only a modest identification loss, CV ~8.5%
  • Retention time SD below 3 s for the 120, 80 and 40 SPD methods, even at low input
  • Identifications evenly distributed across the gradient at every method and load
Evosep EnoWhisper ZoomSingle cellLow input20–120 SPD
PDF · 4 pagesDownload →
Brochure · BR-015B

Evosep Eno sets new standards for routine and high-performance LC-MS

The Evosep Eno release brochure, covering chromatographic performance, reproducibility, sensitivity and quantitative capability against previous standards. Includes comparative HeLa proteomics data, targeted MRM plasma data and column specifications.

  • Peak symmetry improved by up to 35% at 200 samples per day
  • Full width at base improved by more than 30% for the 500 SPD method
  • Retention time reproducibility improved by up to 80% with the 100 SPD method
  • Precursor identifications increased by more than 50% with the faster methods; MRM accuracy above 95%
Evosep EnoBrochureReproducibilityMRM
PDF · 4 pagesDownload →

Evosep Eno Methods

One note per standard and Whisper™ Zoom method — expected chromatography, column and conditions. 10 documents

Method application note · AN-047B

Get started with 500 SPD — Evosep Eno method note

The reference note for the Evosep Eno 500 SPD method: expected high-pressure pump pressure, gradient profile, chromatographic performance and method reproducibility, with the specified column and source-specific heating solutions.

  • 2.3 min MS acquisition, 2.9 min cycle time
  • 4.0 µl/min gradient flow (4.5 µl/min equilibration), EV1182 Performance column at 40 °C
  • FWHM approximately 1.2–1.6 s across the diagnostic peptides
  • Reproducibility shown across three Evosep Eno systems, 15 injections of 50 ng HeLa digest (Pierce)
Evosep EnoMethod note500 SPDEV1182
PDF · 2 pagesDownload →
Method application note · AN-048B

Get started with 300 SPD — Evosep Eno method note

The reference note for the Evosep Eno 300 SPD method: expected high-pressure pump pressure, gradient profile, chromatographic performance and method reproducibility, with the specified column and source-specific heating solutions.

  • 4.0 min MS acquisition, 4.8 min cycle time
  • 4.0 µl/min gradient flow (4.5 µl/min equilibration), EV1182 Performance column at 40 °C
  • FWHM approximately 1.7–2.2 s across the diagnostic peptides
  • Reproducibility shown across three Evosep Eno systems, 15 injections of 50 ng HeLa digest (Pierce)
Evosep EnoMethod note300 SPDEV1182
PDF · 2 pagesDownload →
Method application note · AN-049B

Get started with 200 SPD — Evosep Eno method note

The reference note for the Evosep Eno 200 SPD method: expected high-pressure pump pressure, gradient profile, chromatographic performance and method reproducibility, with the specified column and source-specific heating solutions.

  • 6.4 min MS acquisition, 7.2 min cycle time
  • 2.0 µl/min gradient flow (4.5 µl/min equilibration), EV1182 Performance column at 40 °C
  • FWHM approximately 2.3–3.2 s across the five diagnostic peptides
  • Reproducibility shown across three Evosep Eno systems, 15 injections of 50 ng HeLa digest (Pierce)
Evosep EnoMethod note200 SPDEV1182
PDF · 2 pagesDownload →
Method application note · AN-050B

Get started with 100 SPD — Evosep Eno method note

The reference note for the Evosep Eno 100 SPD method: expected high-pressure pump pressure, gradient profile, chromatographic performance and method reproducibility, with the specified column and source-specific heating solutions.

  • 13.0 min MS acquisition, 14.4 min cycle time
  • 1.0 µl/min gradient flow (2.4 µl/min equilibration), EV1109 Performance column at 40 °C
  • FWHM approximately 2.8–3.4 s across the five diagnostic peptides
  • Reproducibility shown across three Evosep Eno systems, 15 injections of 50 ng HeLa digest (Pierce)
Evosep EnoMethod note100 SPDEV1109
PDF · 2 pagesDownload →
Method application note · AN-051B

Get started with 60 SPD — Evosep Eno method note

The reference note for the Evosep Eno 60 SPD method: expected high-pressure pump pressure, gradient profile, chromatographic performance and method reproducibility, with the specified column and source-specific heating solutions.

  • 22.4 min MS acquisition, 24.0 min cycle time
  • 0.6 µl/min gradient flow (2.4 µl/min equilibration), EV1109 Performance column at 40 °C
  • FWHM approximately 3.8–4.9 s across the diagnostic peptides
  • Reproducibility shown across three Evosep Eno systems, 15 injections of 50 ng HeLa digest (Pierce)
Evosep EnoMethod note60 SPDEV1109
PDF · 2 pagesDownload →
Method application note · AN-052B

Get started with 30 SPD — Evosep Eno method note

The reference note for the Evosep Eno 30 SPD method: expected high-pressure pump pressure, gradient profile, chromatographic performance and method reproducibility, with the specified column and source-specific heating solutions.

  • 45.4 min MS acquisition, 48.0 min cycle time
  • 0.45 µl/min gradient flow, EV1137 Performance column at 40 °C
  • FWHM approximately 5.5–7.4 s across the diagnostic peptides
  • Reproducibility shown across three Evosep Eno systems, 15 injections of 50 ng HeLa digest (Pierce)
Evosep EnoMethod note30 SPDEV1137
PDF · 2 pagesDownload →
Method application note · AN-053B

Get started with Whisper Zoom 120 SPD — Evosep Eno method note

The reference note for the Evosep Eno Whisper Zoom 120 SPD method: expected high-pressure pump pressure, gradient profile, chromatographic performance and method reproducibility, with the specified column and source-specific heating solutions.

  • 10.5 min MS acquisition, 12.0 min cycle time
  • 0.2 µl/min gradient flow (1.0 µl/min equilibration), IonOpticks Aurora Rapid 5×75 at 50 °C
  • FWHM approximately 2.3–2.7 s across the diagnostic peptides
  • Reproducibility shown across three Evosep Eno systems, 15 injections of 5 ng HeLa digest (Pierce)
Evosep EnoMethod note120 SPDWhisper ZoomAurora Rapid
PDF · 2 pagesDownload →
Method application note · AN-054B

Get started with Whisper Zoom 80 SPD — Evosep Eno method note

The reference note for the Evosep Eno Whisper Zoom 80 SPD method: expected high-pressure pump pressure, gradient profile, chromatographic performance and method reproducibility, with the specified column and source-specific heating solutions.

  • 16.3 min MS acquisition, 18.0 min cycle time
  • 0.2 µl/min gradient flow, IonOpticks Aurora Rapid 5×75 at 50 °C
  • FWHM approximately 2.8–3.4 s across the diagnostic peptides
  • Reproducibility shown across three Evosep Eno systems, 15 injections of 5 ng HeLa digest (Pierce)
Evosep EnoMethod note80 SPDWhisper ZoomAurora Rapid
PDF · 2 pagesDownload →
Method application note · AN-055B

Get started with Whisper Zoom 40 SPD — Evosep Eno method note

The reference note for the Evosep Eno Whisper Zoom 40 SPD method: expected high-pressure pump pressure, gradient profile, chromatographic performance and method reproducibility, with the specified column and source-specific heating solutions.

  • 32.8 min MS acquisition, 36.0 min cycle time
  • 0.2 µl/min gradient flow, IonOpticks Aurora Elite 15×75 at 50 °C
  • FWHM approximately 3.2–4.4 s across the diagnostic peptides
  • Reproducibility shown across three Evosep Eno systems, 15 injections of 5 ng HeLa digest (Pierce)
Evosep EnoMethod note40 SPDWhisper ZoomAurora Elite
PDF · 2 pagesDownload →
Method application note · AN-056B

Get started with Whisper Zoom 20 SPD — Evosep Eno method note

The reference note for the Evosep Eno Whisper Zoom 20 SPD method: expected high-pressure pump pressure, gradient profile, chromatographic performance and method reproducibility, with the specified column and source-specific heating solutions.

  • 68.6 min MS acquisition, 72.0 min cycle time
  • 0.2 µl/min gradient flow, IonOpticks Aurora Elite 15×75 at 50 °C
  • FWHM approximately 5.4–7.7 s across the diagnostic peptides
  • Reproducibility shown across three Evosep Eno systems, 15 injections of 5 ng HeLa digest (Pierce)
Evosep EnoMethod note20 SPDWhisper ZoomAurora Elite
PDF · 2 pagesDownload →

Evosep Lupo

Standardized, automated sample preparation with Evokits. 1 document

Application note · AN-058A

Standardized sample preparation at scale with Evokits and Evosep Lupo

Evokit Digest executed on Evosep Lupo and analysed on Evosep Eno, testing transferability between plates, between instruments, and across ten deliberately contaminated sample matrices.

  • 92.1% protein identification overlap between two plates on two separate Evosep Lupo systems
  • Digestion efficiency consistently high at ~99%; protein group CV 12.4–12.7%
  • Stable retention times (SD <0.5 s) in bone tissue spiked with SDS, urea, GdnHCl, Triton X-100 and more
  • Built-in process control and system suitability samples executed as part of the routine workflow
Evosep LupoEvokit DigestSample preparationQCOrbitrap Astral
PDF · 4 pagesDownload →

Evosep One Applications

Applied studies and customer case studies on the Evosep One. 8 documents

Application note · AN-004A

Fast and reproducible phosphoproteomics using MagReSyn Amine and Ti-IMAC HP magnetic beads and the Evosep One

Automated protein aggregation capture digestion and Ti-IMAC HP phosphopeptide enrichment on a KingFisher Flex, combined with Evosep One separation and Orbitrap Exploris 480 detection of HeLa digests. Documents the depth, specificity and replicate reproducibility achievable at 60 samples per day.

  • 7,860 phosphopeptides identified per sample, at 92% enrichment efficiency
  • More than 5,000 localized phosphosites per analysis
  • More than 400 unique phosphopeptides identified per minute of gradient
  • Pearson correlation of 0.97 between replicates, using a 60 SPD method with a 21 minute gradient
PhosphoproteomicsTi-IMACKingFisherOrbitrap Exploris 48060 SPD
PDF · 4 pagesDownload →
Application note · AN-015A

Extended detection of anabolics misuse in high-throughput doping controls with the Evosep One

An Evosep One high-organic method coupled to a Q Exactive HF, screening equine and human urine plus horsehair for anabolic steroid metabolites after enzymatic hydrolysis and mixed-mode extraction. Characterises linearity, detection windows and isomer separation at high throughput.

  • 100 samples per day throughput
  • Linearity assessed at six concentrations spanning 1–100 pg/ml
  • Stanozolol metabolites detectable in urine for up to three months
  • Hair analysis at estimated sub pg/mg concentrations; isomers separated by ~30 seconds
Doping controlAnabolic steroidsUrineHigh organic method100 SPD
PDF · 4 pagesDownload →
Application note · AN-019A

Rapid and robust PTM peptide mapping for biologics with the Evosep One and the SCIEX ZenoTOF 7600

Peptide mapping and post-translational modification analysis on NISTmAb tryptic digest using Evotip Pure with the Evosep One and a SCIEX ZenoTOF 7600, comparing throughput methods and sample loads. Relevant to biopharmaceutical characterisation groups evaluating faster QC-style peptide mapping.

  • Sequence coverage of at least 90% maintained across sample loads and throughputs
  • 100% light chain coverage at 100 ng when fragmentation methods are combined
  • 97.5% heavy chain coverage using the 100 SPD method at 100 ng
  • Median MS1 area CV of 10% across 50 injections; methods evaluated at 60, 100, 200 and 300 SPD
Peptide mappingBiologicsNISTmAbZenoTOF 7600Evotip Pure
PDF · 4 pagesDownload →
Application note · AN-021A

Pushing the boundaries for robust and high-throughput single cell analysis

Single HeLa cells sorted with a cellenONE and analysed on the Evosep One Whisper 40 SPD method coupled to a timsTOF SCP using dia-PASEF. Reports proteome depth per cell, dataset-wide coverage and sample transfer reproducibility.

  • An average of 3,500 protein groups identified per cell
  • Close to 5,500 proteins identified across the 96-cell dataset
  • More than 2,500 proteins significantly regulated between cell size groups
  • 2 × 96 cells prepared within 2.5 hours, using a 15 cm Aurora Elite column
Single cellcellenONEtimsTOF SCPWhisper 40 SPDdia-PASEF
PDF · 4 pagesDownload →
Case study · CS-002A

Unleashing the true power of DIA/SWATH data acquisition with short gradients

ETH Zürich — Aebersold laboratory

The Aebersold laboratory at ETH Zürich, with collaborators at Queen's University Belfast and Northeastern University, applies DIA/SWATH to pre-clinical models and protein complex organisation. They addressed the depth-versus-sample-number trade-off by pairing the Evosep One with short gradients.

  • Analysis cycle time cut from 80 to 24 minutes, raising throughput from 18 to 60 samples per day
  • Approximately 90% of protein detection information retained despite the shorter gradient
  • A mouse liver proteome study completed in about one week instead of two months
  • Complex-centric profiling accelerated roughly tenfold, from 10 days to 1.2 days; 4,065 proteins identified
ETH ZürichDIA/SWATHShort gradientsThroughput
PDF · 4 pagesDownload →
Case study · CS-003A

A fleet of Evosep One instruments supports a proteomics factory at Rapid Novor

Rapid Novor — Kitchener, Ontario

Rapid Novor performs fast-turnaround de novo antibody protein sequencing without genetic information. Rising demand meant eliminating throughput bottlenecks and downtime, so they standardised on eight Evosep One systems fronting eight mass spectrometers.

  • Eight mass spectrometers running 24/7, fronted by eight Evosep One instruments
  • Standard 30 and 60 SPD methods, with switchover between mass spectrometers in under a minute
  • Samples stored on Evotips submerged in liquid for up to a week at room temperature with no losses
  • “The 30 and 60 samples per day methods and integrated Evotip sample clean-up keep our mass spectrometers running at a very high efficiency rate” — Paul Taylor, Core Lab Manager
Rapid NovorDe novo sequencingAntibody sequencingEvotip
PDF · 4 pagesDownload →
Case study · CS-004

Evosep One delivers high throughput and performance consistency in a clinical proteomics lab

Rapid Novor — EasyM assay

Rapid Novor developed EasyM, a serum-based targeted LC-MS assay for monitoring M-protein in multiple myeloma patients as an alternative to bone marrow biopsy. They needed a reproducible, low-maintenance front end able to sustain long unattended sequences.

  • 80% reduction in overhead
  • LC-MS runtime cut 30%, from 30–40 minutes to 22 minutes, doubling throughput
  • 60 runs per day sustained for one to two weeks without intervention
  • “The Evosep One and Evotips eliminate downtime, so we have more time to analyze samples” — Dr Zac McDonald
Rapid NovorClinical proteomicsEasyMMultiple myeloma
PDF · 4 pagesDownload →
Case study · CS-005A

Whisper Zoom delivers sensitivity for highly versatile applications

Uppsala University, Bruker Daltonics and Johns Hopkins University

Researchers working on high-sensitivity proteomics, including single-cell and low-input clinical samples, needed sensitivity gains without sacrificing robustness or standardisation. They adopted Whisper Zoom low-flow methods to achieve it.

  • Phosphoproteomics input material requirement reduced 100-fold
  • 200% increase in peptide identifications with Whisper Zoom 40 SPD versus the standard 30 SPD method
  • Over 1,000 samples run in a single day at 120 SPD, with ~1,500 proteins identified per cell
  • “From day one, we were already connected and collecting single-cell data” — Dr Pierre Sabatier, Uppsala University
Uppsala UniversityWhisper ZoomSingle cellLow input
PDF · 4 pagesDownload →

Evosep One Methods

Standard and specialized separation methods for the Evosep One. 10 documents

Method application note · AN-024A

A standardized separation method with a throughput of 500 SPD

The Evosep One 500 SPD method note: gradient profile, flow conditions, column and emitter options, with chromatographic performance demonstrated on 50 ng tryptic HeLa digest.

  • 2.2 min gradient, 2.9 min cycle time
  • 4 µl/min flow, EV1107 Endurance column at ambient 23 °C
  • FWHM ~1.2–1.5 s across four diagnostic peptides
  • Retention time SD 0.2–0.3 s over ten replicate injections
Evosep OneMethod note500 SPDEV1107
PDF · 2 pagesDownload →
Method application note · AN-005B

A standardized separation method with a throughput of 300 SPD

The Evosep One 300 SPD method note: gradient profile, flow conditions, column and emitter options, with chromatographic performance demonstrated on 50 ng tryptic HeLa digest.

  • 3.2 min gradient, 4.8 min cycle time
  • 4 µl/min flow, EV1107 Endurance column at ambient 23 °C
  • FWHM approximately 1.3–1.6 s
  • Retention time SD 0.4–0.5 s across ten injections
Evosep OneMethod note300 SPDEV1107
PDF · 2 pagesDownload →
Method application note · AN-006B

A standardized separation method with a throughput of 200 SPD

The Evosep One 200 SPD method note: gradient profile, flow conditions, column and emitter options, with chromatographic performance demonstrated on 50 ng tryptic HeLa digest.

  • 5.6 min gradient, 7.2 min cycle time
  • 2 µl/min during the gradient, 4 µl/min for washing and equilibration; EV1107 Endurance at 23 °C
  • FWHM approximately 1.5–2.6 s
  • Retention time SD 0.5–1.2 s across ten replicate injections
Evosep OneMethod note200 SPDEV1107
PDF · 2 pagesDownload →
Method application note · AN-007B

A standardized separation method with a throughput of 100 SPD

The Evosep One 100 SPD method note: gradient profile, flow conditions, column and emitter options, with chromatographic performance demonstrated on 50 ng tryptic HeLa digest.

  • 11.5 min gradient, 14 min cycle time
  • 1.5 µl/min during the gradient, 2 µl/min for equilibration; EV1064 Endurance at 23 °C or EV1109 Performance at 40 °C
  • FWHM approximately 1.8–3.9 s
  • Retention time SD 0.3–1.2 s across ten replicate injections
Evosep OneMethod note100 SPDEV1064EV1109
PDF · 2 pagesDownload →
Method application note · AN-008B

A standardized separation method with a throughput of 60 SPD

The Evosep One 60 SPD method note: gradient profile, flow conditions, column and emitter options, with chromatographic performance demonstrated on 50 ng tryptic HeLa digest.

  • 21 min gradient, 24 min cycle time
  • 1 µl/min during the gradient, 2 µl/min for equilibration; EV1064 Endurance at 23 °C or EV1109 Performance at 40 °C
  • FWHM 2.7–6.1 s depending on peptide and column
  • Retention time SD 0.6–3.1 s across ten injections
Evosep OneMethod note60 SPDEV1064EV1109
PDF · 2 pagesDownload →
Method application note · AN-009C

A standardized separation method with a throughput of 30 SPD

The Evosep One 30 SPD method note: gradient profile, flow conditions, column and emitter options, with chromatographic performance demonstrated on 50 ng tryptic HeLa digest.

  • 44 min gradient, 48 min cycle time
  • 500 nl/min during the gradient, 1500 nl/min for washing; EV1106 Endurance at 23 °C or EV1137 Performance at 40 °C
  • FWHM approximately 4.5–11.0 s depending on column
  • Retention time SD 1.7–4.6 s across eight replicate injections
Evosep OneMethod note30 SPDEV1106EV1137
PDF · 2 pagesDownload →
Method application note · AN-012B

A specialized method — Extended

The Evosep One Extended method note: gradient profile, flow conditions, column and emitter options, with chromatographic performance demonstrated on 50 ng tryptic HeLa digest.

  • 88 min gradient, 93 min cycle time
  • 220 nl/min during the gradient, 1500 nl/min for washing; EV1106 Endurance at 23 °C or EV1137 Performance at 40 °C
  • FWHM 7.5–18.3 s on EV1106 and 7.2–12.9 s on EV1137
  • Retention time SD 1.6–8.7 s (EV1106) and 2.8–7.8 s (EV1137) over eight replicate injections
Evosep OneMethod noteExtendedDeep coverage
PDF · 2 pagesDownload →
Method application note · AN-029A

A specialized method — High Organic

The Evosep One High Organic method note: gradient profile, flow conditions, column and emitter options, with chromatographic performance demonstrated on 50 ng tryptic HeLa digest.

  • 11.5 min gradient, 14 min cycle time
  • 1.5 µl/min during the gradient, 2 µl/min during washing; EV1064 Endurance column
  • FWHM ~2.2–2.5 s across four diagnostic peptides
  • Retention time SD 0.5–0.9 s over ten replicate injections
Evosep OneMethod noteHigh OrganicHydrophobic targetsEV1064
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Application note · AN-011B

Evosep One solves the proteomics dilemma — covering high throughput analysis and proteome depth

An overview of all six standard Evosep One methods from 30 to 500 samples per day, showing how throughput trades against proteome depth on a single platform. Benchmarked on a timsTOF Pro 2 using PAC-prepared tryptic HeLa digest.

  • Gradient and cycle times across the range, from 2.2 / 2.9 min at 500 SPD to 44.0 / 48.0 min at 30 SPD
  • Flow from 4.0 µl/min at 500 SPD down to 0.5 µl/min at 60 and 30 SPD
  • More than 8,000 precursors and just above 2,000 protein groups at 500 SPD
  • Close to 70,000 precursors and more than 7,000 protein groups at 30 SPD from 200 ng HeLa, with protein CV below 10%
Evosep OneMethod overview30–500 SPDtimsTOF Pro 2
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Application note · AN-035C

Democratizing cutting-edge LC-MS performance for high-sensitivity workflows — Whisper Zoom up to 120 SPD

The Whisper Zoom method family for high-sensitivity and low-input proteomics at throughputs up to 120 samples per day. Covers chromatographic performance, load saturation down to single-cell-equivalent inputs, and reproducibility across external laboratories.

  • Whisper Zoom 120 SPD runs a 16.3 min gradient / 18.0 min cycle; 80 SPD 32.5 / 36.0 min; 40 SPD 68.0 / 72.0 min
  • 200 nl/min flow for all Whisper Zoom methods, on IonOpticks Aurora Rapid 5×75 or Aurora Elite 15×75 at 50 °C
  • FWHM 1.6 s at 120 SPD and ~2.1 s at 40 SPD
  • Retention time reproducibility under 5 s intra-instrument and under 16 s inter-instrument across external labs
Evosep OneWhisper Zoom120 SPDHigh sensitivitySingle cell
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Evotip Pure

Carryover, storage, recovery and low-input performance of the Evotip Pure trap column. 3 documents

Application note · AN-022D

Highly efficient disposable trap column with no carry-over

Carryover, quantitative reproducibility, retention time stability and storage recovery for Evotip Pure across a large series of HeLa and plasma digest injections on a ZenoTOF 7600 and a timsTOF Pro 2.

  • Carryover of 0.2% after 50 ng plasma injections and 0.06% after 50 ng HeLa, across 480 injections
  • Average Pearson correlation above 0.99 for 50 ng samples and above 0.97 at 5 ng loads
  • Retention time SD below 1.7 s for five diagnostic peptides across 190 injections
  • Peptides loaded on Evotip can be stored and recovered for up to 28 days without losses
Evotip PureCarryoverReproducibilitySample storageDIA
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Application note · AN-032B

Evotip Pure is the key to accelerate scalable single-cell proteomics

Evotip Pure tested at single-cell-equivalent loads by varying loading volume, adding DDM, measuring carryover from higher-load samples, and tracking stability across a long run series and multi-day on-tip storage.

  • Carryover into 250 pg single-cell-equivalent samples was 0.6% after 50 ng plasma and 0.02% after 50 ng HeLa
  • Carryover in blank injections was 0.2% following both plasma and HeLa samples
  • Median CVs below 7%, with average retention time SD of 2 s for Whisper Zoom 120 SPD
  • More than 90% of precursor identifications preserved after 48 hours on-tip, and close to 75% after 72 hours
Evotip PureSingle cellWhisper ZoomCarryoverSample storage
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Application note · AN-040A

Evotip Pure for end-to-end workflows: a cost-efficient, sustainable and scalable proteomics solution

A fully automated workflow in which HeLa lysate was digested by protein aggregation capture on an AssayMAP Bravo and loaded directly onto Evotip Pure for timsTOF HT analysis, then compared against dried and reconstituted samples.

  • 20% increased recovery through fewer transfer steps
  • As few as 1.25 pipette tips per sample, with reagent use minimised approximately 100-fold
  • Up to 80% utilization of the processed sample, from as little as 1 µg protein input
  • No significant sample loss for up to nine days of storage; direct loading preserved more hydrophobic peptides
Evotip PureAutomationSample preparationSustainabilitydia-PASEF
PDF · 4 pagesDownload →

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