Light extinction and scattering by agarose based resin beads and applications in high-throughput screening
Kittelmann, J. / Hämmerling, F. / Ebeler, M. / Hubbuch, J. (2015)
Journal of Chromatographie A , (2015), 1397, 52-58
- Datum: Juni 2015
Optimization of chromatographic processes by high-throughput screening (HTS) methodologies havebecome a critical part of downstream process development. Nevertheless there are still no non-invasiveoptical methods to characterize resin as well as protein–resin interaction on liquid-handling platformsavailable. Several approaches to automated resin screening in microplates are described in literature,yet all those methods involve indirect measurements by removal of, and sample quantification within,supernatant volumes.
In this work, we introduce light extinction by light scattering to directly assessresin volume and bead density within microplates. Methods for this novel resin characterization aredescribed for 96 and 384-well microplates. An example application demonstrates ligand concentrationmeasurement in microplates with four commercial SP SepharoseTMFast Flow batches.
Further, directquantification of adsorbent bound biomolecules is shown in an example with kinetic protein–resin inter-action measurement in a batch screening process. This new approach is expected to promote batch-basedresin characterization and monitoring on HTS platforms and further miniaturization and increase inthroughput of chromatographic HTS processes.