(W1130-02-08) Room-Temperature Aerosol Dehydration Enables Production of Stable Bevacizumab Powder from FDA-Approved Avastin®

Purpose: Conventional spray drying of monoclonal antibodies typically requires high temperatures >100°C and formulation dilution with additional excipients, which can compromise protein stability and increase manufacturing complexity. This study evaluated the innovative Room Temperature Aerosol Dehydration (RTAD) process [1,2] as a gentle flash dehydration method for producing room-temperature stable bevacizumab powder directly from commercial bevacizumab drug product without requiring dilution or additional excipients beyond the FDA-approved components, and compared the RTAD performance against traditional spray drying.

Methods: Commercial FDA-approved bevacizumab drug formulations (Avastin and its biosimilars, 92 mg/mL total solids content with ~25 mg/mL bevacizumab) were processed as a feedstock using lab-scale RTAD system at 22°C, producing powders at capacity of 50-180 mg/min through flash evaporation of solvent (water). Bevacizumab powder quality was evaluated using VEGF bioassay to assess biological activity and size exclusion chromatography (SEC-HPLC) to quantify dimer formation over 6 months storage at 4°C and 22°C. Particle morphology and size distribution were characterized by scanning electron microscopy. The observed RTAD performance was compared with published spray drying data on bevacizumab [3] obtained at inlet temperatures of ~100°C with outlet temperatures of 50°C and requiring feedstock re-formulation from FDA-approved Avastin to total solid concentrations of 10 mg/mL (~4 mg/mL bevacizumab with ~6 mg/mL excipients) in lab-scale and scale-up studies.

Results: RTAD processing of FDA-approved bevacizumab formulations at 22°C maintained biological activity of powder bevacizumab equivalent to reference standards in VEGF bioassays, with IC50 values remaining unchanged after drying and during 6-month storage at both 4°C and 22°C. SEC-HPLC analysis showed dimer formation remained well below regulatory requirements: 1.4% at both 4°C and 22°C after 6 months storage, meeting target specifications ( < 2%). Scanning electron microscopy demonstrated uniform spherical particle morphology with smooth surfaces, contrasting with the raisin-like or corrugated particles of bevacizumab obtained by the traditional spray drying process [3]. RTAD-formulated bevacizumab powder particles exhibited a consistent median diameter of approximately 2.5 μm, comparable to that achieved by spray drying [3] and potentially suitable for pulmonary delivery. Furthermore, for these spray-drying-like dry particle diameters, RTAD demonstrated improved performance, including usage of feedstock with 9-fold higher total solids concentration (92 mg/mL vs 10 mg/mL) and 6-fold higher API concentration (25 mg/mL vs 4 mg/mL), and up to 3-fold higher powder production rate (180 mg/min vs 60 mg/min). Overall, RTAD enabled direct processing of commercial, FDA-approved bevacizumab drug without dilution or additional excipients, while also reducing the processing temperature dramatically—from 100–120°C in published spray drying studies [3] to just room temperature of 22°C.

Conclusion: Innovative RTAD technology enables gentle dehydration and formulation of dry powders of bevacizumab at room temperature, while preserving bioactivity and maintaining low aggregation levels of bevacizumab during extended room temperature storage. The technology directly processed commercial formulation of monoclonal antibody at significantly higher concentrations without dilution or additional excipients beyond the FDA-approved formulation components, offering substantial manufacturing advantages over conventional spray drying. These new results on bevacizumab and published results on fragment antigen-binding (Fab) region studies [1] demonstrate that the RTAD process represents a promising platform for producing room-temperature stable antibody formulations with reduced processing stress and simplified manufacturing requirements.

References: 1. Poozesh, S.; Mezhericher, M.; Pan, Z.; Chaudhary, U.; Manikwar, P.; Stone, H.A. Rapid Room-Temperature Aerosol Dehydration Versus Spray Drying: A Novel Paradigm in Biopharmaceutical Drying Technologies, Journal of Pharmaceutical Sciences 2024, 113 (4), 974–981.
2. Pan Z, Wang J, Stone HA, Mezhericher M. Room Temperature Aerosol Dehydration of Green Fluorescent Protein. bioRxiv 2024.09.17.613563.
3. Shepard, K. B.; Vodak, D. T.; Kuehl, P. J.; Revelli, D.; Zhou, Y.; Pluntze, A. M.; Adam, M. S.; Oddo, J. C.; Switala, L.; Cape, J. L.; et al. Local Treatment of Non-Small Cell Lung Cancer with a Spray-Dried Bevacizumab Formulation. AAPS PharmSciTech, 2021, 22 (7), 230.

Acknowledgements: This work was supported by the National Science Foundation (NSF) STTR Phase I award No. 2304461 and NSF SBIR Phase II award No. 2451720, and it was also supported by the National Institutes of Health STTR Phase I award No. 1R41TR004571-01A1. The numerical simulations presented in this article were performed on computational resources managed and supported by Princeton Research Computing of the Office of the Dean for Research of Princeton University. Declaration of Competing Interest: H.A.S. and M.M. are co-founders of Inaedis, Inc. and together with Z.P. hold equity in the company.