High-throughput cell quantification assays for use in cell purification development enabling technologies for cell procuction

  • chair:

    Zimmermann, S. / Gretzinger, S. / Scheeder, C. / Schwab, M.L. / Oelmeier, S.A. / Osberghaus, A. / Gottwald, E. / Hubbuch, J. (2016)

  • place:

    Biotechnol J., 2016, 11, 5, 676-686, DOI:10.1002/BIOT.201500577

  • Date: März 2016


High-throughput screening (HTS) technology is gaining increasing importance in downstream process development of cell-based products. The development of such HTS-technologies, however, is highly dependent on the availability of robust, accurate, and sensitive high-throughput cell quantification methods. In this article, we compare state-of-the-art cell quantification methods with focus on their applicability in HTS-platforms for downstream processing of cell-based products. Sensitivity, dynamic range, and precision were evaluated for four methods that differ in their respective mechanism. In addition, we evaluated the performance of these methods over a range of buffer compositions, medium densities, and viscosities, representing conditions found in many downstream processing methods. We found that CellTiter-Glo™ and flow cytometry are excellent tools for high-throughput cell quantification. Both methods have broad working ranges (3–4 log) and performed well over a wide range of buffer compositions. In comparison, CyQuant® Direct and CellTracker™ had smaller working ranges and were more sensitive to changes in buffer composition. For fast and sensitive quantification of a single cell type, CellTiter-Glo™ performed best, while for more complex cell mixtures flow cytometry is the method of choice. Our analysis will facilitate the selection of the most suitable method for a specific application and provides a benchmark for future HTS development in downstream processing of cell-based products.