Single Cell proteomics

Single cell proteomics is a field in rapid development, enabled by significant technological improvements over the last years and it presents a key contribution to answer fundamental biological questions about heterogeneity within complex systems.

Explore how our customers have used the ultra sensitive Evosep One Whisper methods to achieve these technological advantages for single-cell proteomics.

Evotip Pure brings innovative sample storage and introduction for single cell analysis

With single cell proteomics, we can gain valuable biological insights from individual cells, but the limited amount of material in a single cell poses significant challenges. To ensure high-quality, reproducible, and comparable data at a single cell level, standardization is key.

Our Evotip technology is an integral component of the Evosep One system, designed to streamline proteomics workflows. The Evotip enables efficient front-end sample preparation, combining peptide purification with sample loading and elution, facilitating seamless integration with liquid chromatography and mass spectrometry analysis.

Evotip Pure

Capture low-abundance proteins with Whisper Flow Technology

To overcome challenges associated with high-sensitivity analysis, our Whisper Zoom methods combined with fully automated sample preparation on the Evotip Pure is the perfect match when sample amounts are limited.

The plug’n play approach eliminates the need for extensive instrument modifications or specialized expertise. This streamlines your analytical workflow and reduces the barriers to perform high-sensitivity proteomics. And most importantly, the technology seamlessly integrates into standard Evosep One hardware, allowing researchers to adopt the methods quickly and efficiently.

Process thousands of samples on the same emitter and column without loss of performance. The system remains stable over extended periods, ensuring reproducibility for large sample cohorts.

Zoom 120 SPD

Zoom 40 SPD

Zoom 80 SPD

Zoom 20 SPD

The Olsen group

A multi-dimensional view of protein dynamics in single-cell biology

The recent study by the Olsen group at the University of Copenhagen presents groundbreaking advancements in single-cell proteomics, specifically through the use of pSILAC (pulsed Stable Isotope Labeling of Amino Acids in Cell Culture) to examine protein dynamics at the single-cell level.

This research enables an unprecedented view of protein turnover, revealing distinct regulatory mechanisms in dividing versus non-dividing cells. By integrating advanced mass spectrometry and innovative single-cell workflows, the team uncovers subtle differences in protein stability and cellular processes like signal transduction and receptor recycling.

This approach promises significant advancements in understanding cellular heterogeneity, with implications for cancer research, regenerative medicine, and beyond.

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Cellenion CellenOne

Automated Simple, seamless
sample preparation

Sample collection, preparation, and transfer, are often the primary sources of sample losses in high sensitivity proteomics applications.

To address this, a prototype chip, proteoCHIP™ Evo96, compatible with the Cellenion cellenONE instrument has been developed. The chip enables an automated workflow for direct transfer of peptides into Evotips and removes manual handling steps.

This workflow combines sample preparation with the Cellenion platform, pipetting-free transfer of single cell protein digests into Evotips, and analysis with the Whisper 40 SPD method together with the Bruker timsTOF SCP mass spectrometer.

High sensitivity is required in all steps of a workflow and this solution demonstrates excellent proteome coverage for single cells, with up to 4000 protein groups identified per single cell.

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Single cell 2023_Take 4 - The ProteoCHIP

Single cell 2023_Take 5 - Deep visual proteomics

The Mann lab

ultra-high sensitivity workflow from readily available components

In a recent publication from the Mann lab they describe an improved workflow building on recent developments in multiplex-DIA.Their tailored mDIA workflow includes user-friendly dimethyl labelling on the Evotip, automation capabilities, and enhanced quantification options to directly address the needs of many single cell researchers, offering simplicity, reliability, and expanded possibilities for proteomics research.The mDIA workflow, especially when combined with reference channels, offers increased throughput and depth in proteomic analysis. It has the potential to enable the characterization of a significant portion of the expressed proteome in single cells, making it a valuable technology for precision oncology and clinical proteomics.

Their entire ultra-high sensitivity workflow builds entirely upon readily available components. With only three pipetting steps, it is easily automatable on standard liquid handling robots.

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“Single cell proteomics is becoming a practical reality and we are now able to routinely measure large cohorts of single cell proteomes to reveal new biological information. The Evosep One has been pivotal to scale this robustly to thousands of real single cell measurements, making an ideal and standardized platform for future biomarker discovery work”,

Professor Matthias Mann, Max-Planck Institute of Biochemistry,
Proteomics and Signal Transduction

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