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Effect of Buffer Salts on Physical Stability of Lyophilized and Spray-Dried Protein Formulations Containing Bovine Serum Albumin and Trehalose

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Abstract

This study examined the effect of buffer salts on the physical stability of spray-dried and lyophilized formulations of a model protein, bovine serum albumin (BSA). BSA formulations with various buffers were dried by either lyophilization or spray drying. The protein powders were then characterized using solid-state Fourier transform infrared spectroscopy (ssFTIR), powder X-ray diffraction (PXRD), size exclusion chromatography (SEC), solid-state hydrogen/deuterium exchange with mass spectrometry (ssHDX-MS), and solid-state nuclear magnetic resonance spectroscopy (ssNMR). Particle characterizations such as Brunauer-Emmett-Teller (BET) surface area, particle size distribution, and particle morphology were also performed. Results from conventional techniques such as ssFTIR did not exhibit correlations with the physical stability of studied formulations. Deconvoluted peak areas of deuterated samples from the ssHDX-MS study showed a satisfactory correlation with the loss of the monomeric peak area measured by SEC (R2 of 0.8722 for spray-dried formulations and 0.8428 for lyophilized formulations) in the 90-day accelerated stability study conducted at 40°C. mDSC and PXRD was unable to measure phase separation in the samples right after drying. In contrast, ssNMR successfully detected the occurrence of phase separation between the succinic buffer component and protein in the lyophilized formulation, which results in a distribution of microenvironmental acidity and the subsequent loss of long-term stability. Moreover, our results suggested that buffer salts have less impact on physical stability for the spray-dried formulations than the lyophilized solids.

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ACKNOWLEDGMENTS AND DISCLOSURES

Qi (Tony) Zhou and Tarun Tejasvi Mutukuri were supported by the National Institute of Allergy and Infectious Diseases of the National Institute of Health under Award Number R01AI146160. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institute of Health. Yong Du appreciates MRL postdoctoral research program. The authors are grateful for the helpful discussion and MS access from Dr. Elizabeth M. Topp. The authors have no conflict of interest in this work.

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Authors and Affiliations

  1. Department of Industrial and Physical Pharmacy, College of Pharmacy, Purdue University, 575 Stadium Mall Drive, West Lafayette, Indiana, 47907, USA

    Tarun Tejasvi Mutukuri & Qi Tony Zhou

  2. Discovery Pharmaceutical Sciences, Merck & Co., Inc., South San Francisco, California, 94080, USA

    Jing Ling

  3. Analytical Research and Development, Merck & Co., Inc., Rahway, New Jersey, 07065, USA

    Yong Du & Yongchao Su

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  1. Tarun Tejasvi Mutukuri
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Mutukuri, T.T., Ling, J., Du, Y. et al. Effect of Buffer Salts on Physical Stability of Lyophilized and Spray-Dried Protein Formulations Containing Bovine Serum Albumin and Trehalose. Pharm Res 40, 1355–1371 (2023). https://doi.org/10.1007/s11095-022-03318-7

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