Acquisition of a cellenONE F1.4

Novo Nordisk Foundation Center for Protein Research (CPR) was established in 2007 at the Faculty of Health and Medical Sciences, University of Copenhagen, to promote basic and applied discovery research on human proteins of medical relevance. The Center comprises a wide range of expertise and skills within its research programs spanning disease systems biology, proteomics, high throughput protein production and characterization, chemical biology, disease biology, and protein therapeutics. CPR contributes to the progress of translational research within medicine and provides fundamental insights which can be used to promote drug discovery and development. CPR’s vision is to become a world-leader in exploring how protein modifications and their functional networks drive fundamental biological processes that underlie health and disease. It has the mission to integrate innovative protein technologies, big data analytics and mechanism-based research to advance understanding of disease-relat

High sensitivity sample preparation is the key for successful single-cell proteomics. The cellenONE® F1.4 includes a Single Cell Isolation, Sorting, and Dispensing System. Cells are dispensed in tiny, sub-nanoliter droplets of adjustable volume, enabling extreme assay miniaturization and corresponding dramatic savings in the cost of reagents. Droplets are dispensed with microarray-type accuracy and precision with typical spot pitches of down to 100 microns in any pattern and into 96-384- or 1536-well plates or nanowell vessels or onto slides or custom substrates. The cellenONE® is available with a second dispensing channel to permit additional on-board proteomics sample preparation, temperature control for target substrates, as well as humidity and dewpoint control to further enhance sample integrity and prevent evaporation. In addition to high-resolution bright field microscopy mode, cells can also be interrogated by fluorescence in four distinct channels- green, red, orange, and blue. The gentle, piezo-driven acoustic dispense technology preserves the viability of dispensed cells. Cells of 2 – 80 microns in diameter can be detected in bright field mode, while cells less than 1 micron can be detected using fluorescent mode. Images of the droplet containing single, isolated cells and of the cells as they proceed toward the dispensed droplet portion of the nozzle are available for real-time observation and are also preserved for documentation of singularity and review of morphological characteristics. Only a few microliters of sample are needed, and recovery rates for rare cells has been documented to exceed 90%, with no sample wastages, as cells that are not isolated can be recovered and re-processed. Proteomics sample preparation including protein extraction, protease-digestion, and TMTpro 16-plex-labeling has been integrated in a streamlined workflow making this system the only one to enable automated highly sensitive single-cell proteomics sample preparation. We plan to use the cellenONE for single-cell proteomics sample preparation for ultra-sensitive LC-MS/MS analysis with specific applications to study cellular heterogeneity in human tissues in health and disease.

The cellenONE® F1.4 instrument is a so-called unika instrument as no other vendors have an instrument on the market that can do single-cell sorting and highly sensitive nL single-cell proteomics sample preparation in one integrated and automated workflow.

The cellenONE F1.4 device utilizes, exclusively, a collection of intellectual property, including that described in the following patents:

Method and apparatus for single particle deposition (EP16000699, US20170274689A1)

Method and dispenser device for depositing a substance on a target substrate (WO2015127945A1, US20170072417A1)

Method and devices for depositing samples on an electrically shielded substrate (EP1675681B1, US8614100B2)

Spotting plate and process for its production (EP2636453A3, US20130237443A1)

Microdispenser and associated operating method (EP1886146B1, US20120312836A1)

Microdispenser and associated operating method (WO2006128662A1, US8273307B2)

Single-cell proteomics based on mass spectrometric analysis of protein digests from single cells is a new and emerging field in biomedicine. The cellenONE F1.4 is the only commercially available instrument, which can automatically do Single Cell Isolation, Sorting, and Dispensing in precise nL volumes and combine this with an integrated ultrasensitive single-cell proteomics preparation workflow involving protein extraction, protease-digestion, and TMTpro-16-plex labeling of individual single cell channels and carrier proteome booster channel.