Category: Automation and High-Throughput Technologies
In-Process analytical testing is a critical component in the protein biologics pipeline; essential for monitoring product quality to provide necessary information at process decision points. High-throughput and reliable data generation enables rapid and accurate in-process assessment. Conventional analytical methods for in-process testing of therapeutic antibodies can be labor and resource intensive. Towards this goal, we have developed and implemented automated workflows for antibody purification and oligosaccharide profiling sample preparation on the Tecan Fluent liquid handling platform. Antibodies are purified from clarified cell culture harvest using Protein A CentriColumns®; purification of 96 samples can be accomplished in ~ 2hrs and sufficient antibody is obtained for analytical measurements of product quality attributes.
Antibody purification method was evaluated for reproducibility and robustness using clarified cell culture harvests from 2 different antibody molecules. Four different clone harvests with mass loads of 600 - 2000μg gave yields of ~95% with CV of less than 10. Column cycling shows the columns can be re-used up to 8 cycles with good reproducibility of yield (~90%). Automated glycan sample preparation method transfers the commercially available Waters GlycoWorks™ RapiFluor™ N-Glycan Kit single tube manual method to 96-well plate format. The RapiFluor™ N-glycan method was evaluated with increased reagent volumes and incubation times for the denaturation, deglycosylation and labeling steps. Manual and automated glycan sample preparation methods were comparable with no significant difference in the peak area percentages of the quantitated glycan species.
Sharmila Sivendran– Scientist, Sanofi Genzyme, Framingham, MA
Sharmila Sivendran is a Scientist in BioProcess Analytics within BioAnalytics in BioPharmaceutics Development. Prior to joining Sanofi-Genzyme, Sharmila worked at GlaxoSmithKline and Mount Sinai Medical Center in the Assay Development and High-throughput Screening Core Facilities. Sharmila received her Ph.D. in Biochemistry from University of Delaware, focusing on biochemical and biophysical characterization of proteins. Her postdoctoral work at the University of Pennsylvania in the Department of Chemical and Biomolecular Engineering focused on automation and development of miniaturized biochemical and cell-based assays for high-throughput screens.