The whisper 80 SPD method
High throughput single cell proteomics
Single cell proteomics is now at a place, where a proteome depth of a few thousands proteins is achievable on a routine basis with the Evosep One. This leaves opportunities to scale up the sampling rate for improved statistics.
The Whisper 80 SPD method is specifically designed to scale this field for high sensitivity work and facilitates a 15 min gradient running with a flowrate of 100 nl/min.
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.
HIGHER THROUGHPUT AND EXCELLENT SENSITIVITY WITH WHISPER 80 SPD
The sensitivity of the Whisper 80 samples per day method was evaluated using the Aurora Rapid 75 column with a dilution series of HeLa tryptic peptides analyzed in eight replicates, ranging from low loads of 62.5 pg up to saturating levels of 32 ng peptide material.
A timsTOF SCP mass spectrometer using data-independent acquisition parallel accumulation serial fragmentation (dia-PASEF) and analyzed with DIA-NN using a predicted library was used to generate this dilution series.
Speeding up Single Cell Analysis
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.
The 80 SPD method identified more than a thousands proteins with this workflow, and has has interesting perspectives for scalable workflows using multiplexing strategies.
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