Microwell Plate-Based Dynamic Light Scattering as a High-Throughput Characterization Tool in Biopharmaceutical...

High-throughput light scattering instruments are widely used in screening of biopharmaceutical formulations and can be easily incorporated into processes by utilizing multi-well plate formats. High-throughput plate readers are helpful tools to assess the aggregation tendency and colloidal stability of biological drug candidates based on the diffusion self-interaction parameter (k). However, plate readers evoke issues about the precision and variability of determined data. In this article, we report about the statistical evaluation of intra- and inter-plate variability (384-well plates) for the k analysis of protein and peptide solutions. ANOVA revealed no significant differences between the runs. In conclusion, the reliability and precision of k was dependent on the plate position of the sample replicates and k value. Positive k values (57.0 mL/g, coefficients of variation (CV) 8.9%) showed a lower variability compared to negative k values (-14.8 mL/g, CV 13.4%). The variability of k was not reduced using more data points (120 vs. 30). A k analysis exclusively based on center wells showed a lower CV (<2%) compared to edge wells (5-12%) or a combination of edge and center wells (2-5%). We present plate designs for k analysis within the early formulation development, screening up to 20 formulations consuming less than 50 mg of active pharmaceutical ingredient (API).