title: Robust high-throughput batch screening method in 384-well format with optical in-line resin quantification.
abstract: High-throughput batch screening technologies have become an important tool in downstream process development. Although continuative miniaturization saves time and sample consumption, there is yet no screening process described in the 384-well microplate format. Several processes are established in the 96-well dimension to investigate protein-adsorbent interactions, utilizing between 6.8 and 50 μL resin per well. However, as sample consumption scales with resin volumes and throughput scales with experiments per microplate, they are limited in costs and saved time. In this work, a new method for in-well resin quantification by optical means, applicable in the 384-well format, and resin volumes as small as 0.1 μL is introduced. A HTS batch isotherm process is described, utilizing this new method in combination with optical sample volume quantification for screening of isotherm parameters in 384-well microplates. Results are qualified by confidence bounds determined by bootstrap analysis and a comprehensive Monte Carlo study of error propagation. This new approach opens the door to a variety of screening processes in the 384-well format on HTS stations, higher quality screening data and an increase in throughput.
authors.affiliation: Section IV: Biomolecular Separation Engineering, Institute of Engineering in Life Sciences, Karlsruhe Institute of Technology (KIT), Engler-Bunte-Ring 1, 76131 Karlsruhe, Germany.
authors.forename: Jörg
authors.initials: J
authors.lastname: Kittelmann
authors.affiliation: Bioprocess Engineering Group (BPE), Department of Biotechnology, Delft University of Technology, Julianalaan 67, 2628 BC Delft, The Netherlands.
authors.forename: Marcel
authors.initials: M
authors.lastname: Ottens
authors.affiliation: Section IV: Biomolecular Separation Engineering, Institute of Engineering in Life Sciences, Karlsruhe Institute of Technology (KIT), Engler-Bunte-Ring 1, 76131 Karlsruhe, Germany. Electronic address: Juergen.Hubbuch@kit.edu.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Hubbuch
mesh_terms: D002853:Chromatography, Liquid / Q000295:instrumentation* / Q000379:methods*; D016000:Cluster Analysis; D004867:Equipment Design; D057166:High-Throughput Screening Assays / Q000379:methods*; D055616:Microtechnology / Q000295:instrumentation*; D009010:Monte Carlo Method; D009113:Muramidase; D011108:Polymers; D015203:Reproducibility of Results; D012685:Sepharose
chemical_list: D011108:Polymers; C089543:Toyopearl Phenyl 650 S; D012685:Sepharose; D009113:Muramidase
keywords: Batch isotherm; Bootstrap analysis; High-throughput screening (HTS); Liquid handling station (LHS); Monte Carlo; Resin quantification