The whisper 40 SPD method

EVOSEP+ METHOD

ROBUST AND HIGH-THROUGHPUT SINGLE CELL ANALYSIS

Whisper 40 SPD beta is specifically developed for single cell analysis, where one of the main challenges is to maintain sensitivity and robustness throughout the workflow.

It runs with a gradient flow of 100 nl/min with a 31 minutes method duration to accommodate the sensitivity and speed needed for optimal separation of minute amount of material.

IT’S ALL IN THE EVOTIP

The Evotip, a disposable trap column, is the core of our technology. The Evotip serves as an integrated purification step, where contaminants are left on the tip, and analytically relevant peptides are eluted.

The Evotip has proven to be extraordinary important for ultra-sensitive analysis, as very small sample amounts can be loaded in higher volumes.

Furthermore, loaded Evotips can be stored cold until analysis, which is especially important for single cell proteomics, where very limited sample amount sets higher demands to the storage of peptide and become a crucial factor for reproducible analysis.

EXCELLENT COVERAGE WITH UP TO 2500 PROTEINS IDENTIFIED FROM A SINGLE CELL

Robust and scalable workflows are needed to gain biological insight from single cell analysis. The cellenONE represents a unique technology allowing high-throughput image-based automated sorting and dispensing of cells.

In combination with the Aurora Elite column from IonOpticks, Whisper methods on the Evosep One and dia-PASEF on the timsTOF SCP, the pipeline provides an excellent workflow for robust and high-throughput single cell analysis at competitive sensitivity.

This workflow maps single cell proteomes with a coverage of up to 2,500 proteins, pushing the boundaries of protein-level insight achievable across heterogeneous cell populations.
Read more about the data here

HIGH PERFORMANCE POWERED BY AURORA ELITE COLUMNS

The Whisper 40 SPD method is designed for our EV1112 column and EV1111 fused silica emitter. For competitive performance, we recommend to use the Aurora Elite column from IonOpticks. This can be used directly for the Whisper 40 SPD and 20 SPD methods, while maintaining the column temperature at 50 °C.

Combined with the latest generation Evotip Pure, the chromatography enables a median FWHM of 3 seconds of all identified precursors, based on 5 ng HeLa.

Three diagnostic peptides eluting across the gradient depict specific retention times and peak properties on the figure.

SuPERIOR REPRODUCIBILITY WITH DILUTION SERIES

Excellent chromatography with sharp symmetric peaks serves as the foundation to employ the sensitivity and speed on modern mass spectrometers.

In collaboration with Bruker, we evaluated the sensitivity of our workflow with a dilution series of HeLa tryptic peptides analyzed in hexuplicate (Pierce), ranging from low loads of 62.5 pg up to saturating levels of 32 ng peptide material using dia-PASEF on a timsTOF SCP mass spectrometer.
This led to 2,000 proteins from 250 pg, whereas the high load of 32 ng resulted in almost 7,500 proteins.

The dilution series revealed excellent linearity of identifications as well as reproducibility across the six injections.

Get well started with Whisper

We would like to get you started well with the Whisper methods. Here you can find a step-by-step guide describing how to connect your column and emitter to obtain optimal peak shapes and thereby the best chromatographic performance.

Download guide

Webinar

Whisper is here

Learn more about the advantages of the pioneering Whisper Flow Technology in this introduction by Nicolai Bache, Head of Applications at Evosep. 

Watch full webinar on demand here

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Whisper is part of our dynamic beta environment, where updates are to be expected. Sign up here to get notified with relevant information.

explore all available methods with evosep+

Find the method fitting your research need and get inspired with data from Evosep users.

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This EVOSEP+ method enables small-molecules analysis with significantly more hydrophobic targets.

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Whisper
40 SPD beta

EVOSEP+ method

Ultra-sensitive method ideal for single-cell analysis.

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20 SPD beta

EVOSEP+ method

A longer method for robust and sensitive applications.

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ULTRA-SENSITIVE AND STABLE PERFORMANCE WITH WHIsPER FLOW TECHNOLOGY
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Whisper™ step-by-step guide
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Column ovens for Evosep performance columns
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columns and emmitters overview
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EVOSEP ONE: A GRADIENT OFF-SET FOCUSING UHPLC INSTRUMENT FOR ROBUST AND HIGH THROUGHPUT PROTEOMICS

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