Design and Optimization of a Temperature-Stable Dry Powder BCG Vaccine
Loss of vaccine potency due to extreme temperature exposure during storage and transport remains a significant obstacle to the success of many vaccines, including the Bacille Calmette-Guérin (BCG) vaccine, the only vaccine available against Mycobacterium tuberculosis. BCG is a live, attenuated vaccine requiring refrigerated storage for viability. In this study, we formulated a temperature-stable BCG dry powder using the spray drying technique.
We employed a factorial design to optimize our formulation of stabilizing excipients that included L-leucine, bovine serum albumin, polyvinylpyrrolidone, mannitol, and trehalose. Powders were characterized for their particle size, yield, water retention and uptake, glass transition temperature, and aerosol performance. Three optimal powder carrier mixtures were selected from the factorial design for BCG incorporation based on their stability-promoting and powder flow characteristics. Vaccine powders were also assessed for BCG viability and in vivo immunogenicity after long-term storage.
Live BCG was successfully spray-dried using the optimized carriers. Dry powder BCG showed no loss in viability (25°C, up to 60% relative humidity; RH) and ~2-log loss in viability (40°C, 75% RH) after one year of storage. The aerodynamic size of the powders was in the respirable range. Further, when healthy mice were immunized intradermally with reconstituted BCG powders (storage for 2 years), the vaccine retained its immunogenicity.
We developed a spray-dried BCG vaccine that was viable and antigenic after long-term storage. To our knowledge, this is a first study to show room temperature stability of live BCG vaccine without any loss in viability for 12 months.
Key WordsBCG cold-chain factorial design live bacterial vaccine pulmonary vaccination spray drying tuberculosis vaccine formulation vaccine stability
Acknowledgements and Disclosure
The authors would like to acknowledge Dr. Avni Patel for her help with the initial spray drying experiments. Laura Stephens for her help with the phagocytosis assay. We would also like to thank Dr. Hien Pham for the use of SEM, Dr. Eric Peterson for XRD, and Dr. Stephen Jett for TEM analysis.The authors declare no conflict of interest.
This work was funded in part by the Bill and Melinda Gates Grand Challenges Exploration (OPP1061393; PI- Dr. Pavan Muttil), National Institute of Allergy and Infectious Diseases (T32 Training Grant No: A1007538; Recipient- DNP), and by the National Research Foundation (Grant# 105830, South Africa; PI- Dr. Bernard Fourie).
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