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The Bioprocessing Summit convenes more than 1,500 international bioprocess professionals to share practical solutions for today's bioprocess challenges. Now in its eleventh year, the event has grown to include 16 distinct meetings with weeklong programming on upstream and downstream processing, analytical development and quality, formulation and stability, cell and gene therapy production, and manufacturing. Along with the impressive array of conferences, the Summit also includes short courses and training seminars that provide in-depth coverage of critical bioprocess topics.
High Throughput Sialylation Measurement on Octet Label Free Instrument for Cell Line Development
Presenter: Hongshan Li
Co-Authors: Danfeng Yao; Spence Borg; Sriram Kumaraswamy
A high-throughput method for relative screening of terminal sialic acid content was developed on an Octet platform to expedite cell line development. The method is based on the use of ForteBio’s Sialic Acid (GlyS) kit to bind sialic acid on glycoproteins. This method can specifically screen the sialylation levels of secreted proteins in crude cell culture samples and does not require purified samples. Using this method, 96 crude cell culture samples can be screened for sialylation in 60 minutes or less. In a comparison study between the Octet GlyS kit method and HPLC, a linear correlation (R2 = 0.9) was observed between the GlyS kit binding signal (nm/ug) obtained using an Octet HTX system and the total sialic acid content measured by HPLC. The samples ranged between 1-25 mol of total sialic acid per mol of protein. This method can screen relative sialylation levels in crude samples in a high through-put manner and can be used to monitor quality attributes early in cell line development for therapeutic proteins hence accelerating the process.
Combining Single-Cell Isolation and Microplate Imaging to Increase Colony Outgrowth Efficiency with Higher Assurance of Clonality
Presenter: Hongshan Li
Author: Shan Liu
The CloneSelect™ Single-Cell Printer™ Series addresses limitations in current cell line development workflows by offering image-based evidence for clonality, fast and efficient single-cell deposition based on brightfield (c. sight™ and f. sight™ systems) or optional fluorescence (f. sight system only), excellent post-deposit cell growth, and minimal risk of cross-contamination. When coupled with the rapid imaging technology of the CloneSelect Imager (CSI), this platform ensures a significant increase in clonal outgrowth and provides assurance of monoclonality. Here, we demonstrate an optimized workflow that combines single-cell deposition and monoclonality verification. We used the f. sight system to deposit single cells into a microplate with image-based evidence of clonality and the CSI for additional confirmation of the presence of a single cell per well and for monitoring cell growth over several days.