Bruker Corporation and Evosep have announced major progress in high-sensitivity, quantitative true single-cell proteomics, using a modified timsTOF Pro mass spectrometer connected to an Evosep One low-flow chromatography system.
These advances have been demonstrated in a paper on Bioarchive. Key results include the first untargeted and unbiased analysis of true single cells achieving an identification depth of up to 1500 protein groups from single FACS-sorted and individually digested HeLa cells. This sensitivity was achieved by the combined Evosep–timsTOF platform using crucial innovations.
Two new ultralow-flow methods on an otherwise unmodified Evosep One system offer 100 nL/min gradients that lead to approximately 10 times higher sensitivity than gradients at 1 µL/min, confirming theoretical expectations. These new nano-flow methods, using the Whisper flow technology, have the same low overhead as the other established Evosep One methods, and offer a throughput of either 20 or 40 samples per day. They make single-cell analysis robust, as shown by the analysis of 420 single cells that were analysed on the same column. The workflow retrieved known and unexpected proteins, changing abundance upon chemically arresting the cell cycle at discrete points.