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Easy and Accessible Workflow for Label-Free Single-Cell Proteomics

Updated: Sep 28, 2023




Ryan Kelly's Research Group has unveiled a accessible workflow to simplify single-cell proteomics (SCP). Their recent publication in the Journal of the American Society for Mass Spectrometry sheds light on the significance of SCP in revealing unparalleled insights that aren't possible with bulk-scale protein measurements or other single-cell profiling methods.

Historically, widespread adoption of SCP was hindered due to the need for specialized equipment, consumables, and expertise. Moreover, while commercial platforms emerged for SCP, their high costs and technical complexity remained a barrier for many labs.

However, the tide is turning. Kelly's team tested the HP D100 Single Cell Dispenser for label-free SCP, marking it as a game-changer. Notably:

  • Affordability & Precision: The HP D100 is cost-effective and showcases remarkable precision, dispensing reagents from 200 nL to 2 μL.

  • Efficiency: Utilizing the HP D100, single cells were isolated and prepared for SCP within 384-well PCR plates. A staggering ~97% of these wells delivered the expected proteome coverage of a single cell when proper procedures were followed.

  • Simplification without Compromise: This dispenser, coupled with a one-step sample processing, offers a swift and user-friendly SCP workflow. Moreover, it does so without compromising proteome coverage, and is compatible with standard commercial well plates, facilitating seamless auto-sampling.

  • Analysis Flexibility: For the analysis, both home-packed 30 μm i.d. nanoLC columns and off-the-shelf 50 μm i.d. columns were used. The latter identified approximately 35% fewer proteins but, when combined with the well plate-based platform, presents a commercially viable and budget-friendly option for SCP sample preparation and separation.

This innovative workflow is expected to democratize access to SCP, empowering more laboratories to explore the intricacies of individual cells.


Please reach out to Kelly's group for any question or comments, https://chembio.byu.edu/directory/ryan-kelly

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