Screening Developability and Pre-Formulation of Biotherapeutics with High-Throughput Dynamic Light Scattering (HT–DLS)

Significance of the Topic

Early evaluation of biotherapeutic developability is crucial to reduce costly downstream failures. High-throughput dynamic light scattering (HT–DLS) offers rapid, low-volume, noninvasive measurements of protein aggregation, stability, and viscosity, supporting quality-by-design and formulation screening.

Objectives and Study Overview

This white paper presents the capabilities of Wyatt Technology’s DynaPro Plate Reader (DPR) to perform automated HT–DLS in multiwell plates, enabling screening of diverse formulation and environmental conditions for biotherapeutic candidates.

Methodology and Instrumentation Used

The DPR integrates dynamic light scattering (DLS), static light scattering (SLS), onboard imaging, and 21CFR11-compliant DYNAMICS SP software. Using standard 96- and 384-well plates, it performs unattended measurements of hydrodynamic radius, molar mass, colloidal interaction parameters (kD and A2), thermal and chemical stability via temperature ramps and denaturant gradients, and viscosity via polystyrene probe beads.

Main Results and Discussion

• Gross aggregation screening demonstrated rapid quantification of submicron aggregates in lyophilized monoclonal antibody formulations, visualized with intensity distributions and heat maps.
• Onboard imaging identified precipitation and contaminants post-analysis.
• Thermal stability assays characterized unfolding and aggregation transitions of lysozyme and IgG, distinguishing onset temperatures for conformational changes and aggregate formation using DLS and SLS.
• Chemical denaturation studies provided direct, reliable unfolding data without fluorophore interference.
• Colloidal stability analysis determined interaction parameter kD across pH values for multiple proteins, predicting aggregation propensity.
• Viscosity measurements correlated DLS-derived viscosities with traditional rheometry, enabling formulation screening for injectability.

Benefits and Practical Applications

HT–DLS accelerates developability assessment by delivering 10–100× more measurements in minutes, reducing sample volume and operator intervention. It supports DoE and QbD strategies, improves data quality, and is suitable for GLP/GMP labs concentrating on candidate selection, stability profiling, and formulation optimization.

Future Trends and Applications

Integration with fully automated workflows, real‐time process analytical technology (PAT), and machine learning for predictive analytics will expand HT–DLS utility. Emerging applications include antibody–drug conjugates, viral vectors, and high‐throughput screening of complex biologics.

Conclusion

Automated HT–DLS on the DynaPro Plate Reader provides comprehensive, high‐throughput insights into biotherapeutic developability, enabling informed early‐stage selection and robust formulation strategies.

Instrumentation Used

• DynaPro Plate Reader (DPR)
• DYNAMICS SP software (21CFR11 compliant)
• Standard 96‐ and 384‐well microplates

References

1. Rader R. FDA Biopharmaceutical Product Approvals and Trends in 2012. BioProcess International. 2013.
2. Some D. Biopharmaceutical Candidate Screening with Automated Dynamic Light Scattering. Chromatography Online. 2015.
3. Cockrell GM, Wolfe MS, Wolfe JL, Schöneich C. Photoinduced aggregation of a model antibody‐drug conjugate. Mol. Pharm. 2015;12:1784–1797.
4. Yu Z, Reid JC, Yang YP. Utilizing dynamic light scattering as a process analytical technology for protein folding and aggregation monitoring in vaccine manufacturing. J. Pharm. Sci. 2013;102:4284–4290.
5. Menzen T, Friess W. Temperature‐ramped studies on the aggregation, unfolding, and interaction of a therapeutic monoclonal antibody. J. Pharm. Sci. 2014;103:445–455.