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TECHNICAL NOTE Sizing accuracy and intra-assay precision of DLS measurements with Prometheus Panta Introduction Prometheus Panta uses nano differential scanning fluorimetry (nanoDSF), dynamic light scattering (DLS), and backreflection for high-resolution, multi-parameter stability characterization. The simultaneous collection of data on thermal unfolding, particle sizing, and aggregation throughout the entire thermal ramp allows for the identification of the temperature at which the protein unfolds, forms amorphous aggregates, or even begins to change size or polydispersity. Scientists in the field of drug discovery, particularly biologics, use DLS to make critical decisions about drug candidate stability. This creates a need for instruments that exhibit strong sizing accuracy and precision. These properties can be evaluated using standards and references from the National Institute of Standards and Technology (NIST). Here, the NIST monoclonal antibody (NISTmAb) reference material (RM) RM8671, Lysozyme, and a set of monodisperse particle size standards were used to test the sizing accuracy and intra-assay precision of Prometheus Panta's DLS optical module. Results Sizing accuracy The sizing accuracy was assessed by measuring the hydrodynamic radii (r H ) of different particle size standards from Thermo Fisher Scientific that ranged in diameter from approximately 20 nm to 1 µm. Prior to DLS measurement, each size standard was sonicated for 60 seconds and, in case of the 20, 60, and 100 nm particles, diluted 1:100 in Philipp Schramm, Silvia Würtenberger, Christian Kleusch NanoTemper Technologies GmbH, Munich, Germany

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