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

• Main Authors: Katharina Dauer (Author), Stefania Pfeiffer-Marek (Author), Walter Kamm (Author), Karl G. Wagner (Author)
• Format: Book
• Published: MDPI AG, 2021-01-01T00:00:00Z.
• Subjects: article
• Online Access: Connect to this object online.

Summary

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 (<i>k_D</i>). 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 <i>k_D</i> analysis of protein and peptide solutions. ANOVA revealed no significant differences between the runs. In conclusion, the reliability and precision of <i>k_D</i> was dependent on the plate position of the sample replicates and <i>k_D</i> value. Positive <i>k_D</i> values (57.0 mL/g, coefficients of variation (<i>CV</i>) 8.9%) showed a lower variability compared to negative <i>k_D</i> values (−14.8 mL/g, <i>CV</i> 13.4%). The variability of <i>k_D</i> was not reduced using more data points (120 vs. 30). A <i>k_D</i> analysis exclusively based on center wells showed a lower <i>CV</i> (<2%) compared to edge wells (5-12%) or a combination of edge and center wells (2-5%). We present plate designs for <i>k_D</i> analysis within the early formulation development, screening up to 20 formulations consuming less than 50 mg of active pharmaceutical ingredient (API).

Item Description

10.3390/pharmaceutics13020172
1999-4923