Lyophilization and spray drying are widely used to manufacture solid forms of therapeutic proteins. Lyophilization is used to stabilize proteins vulnerable to degradation in solution, whereas spray drying is mainly used to prepare inhalation powders or as an alternative to freezing for storing bulk drug substance. Both processes impose stresses that may adversely affect protein structure, stability and bioactivity. Here, we compared lyophilization with and without controlled ice nucleation, and spray drying for their effects on the solid-state conformation and matrix interactions of a model IgG1 monoclonal antibody (mAb).
Solid-state conformation and matrix interactions of the mAb were probed using solid-state hydrogen-deuterium exchange with mass spectrometric analysis (ssHDX-MS), and solid-state Fourier transform infrared (ssFTIR) and solid-state fluorescence spectroscopies.
mAb conformation and/or matrix interactions were most perturbed in mannitol-containing samples and the distribution of states was more heterogeneous in sucrose and trehalose samples that were spray dried.
The findings demonstrate the sensitivity of ssHDX-MS to changes weakly indicated by spectroscopic methods, and support the broader use of ssHDX-MS to probe formulation and process effects on proteins in solid samples.
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Attenuated total reflectance
Bovine serum albumin
Controlled ice nucleation
Fourier transform infrared
Hydrogen deuterium exchange with mass spectrometric analysis
- IgG :
- Kfast :
Rate of exchange of amide hydrogen atoms in the fast exchanging pool
- Kslow :
Rate of exchange of amide hydrogen atoms in the slow exchanging pool
- Nfast :
Number of amide hydrogen atoms in the fast exchanging pool
- Nslow :
Number of amide hydrogen atoms in the slow exchanging pool
Powder x-ray diffraction
Scanning electron microscope
Solid-state; indicates that an analytical method has been applied to a solid sample
- ssPL-MS :
Solid-state photolytic labeling with mass spectrometric analysis
Uncontrolled ice nucleation
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ACKNOWLEDGMENTS AND DISCLOSURES
The IgG1 mAb used in this study was provided by Pfizer, Inc. (Chesterfield, MO). This work was supported in part by a Purdue Research foundation grant and a McKeehan Graduate Fellowship awarded by Purdue University to EM.
Guest Editors: Tony Zhou and Tonglei Li
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Moussa, E.M., Wilson, N.E., Zhou, Q.T. et al. Effects of Drying Process on an IgG1 Monoclonal Antibody Using Solid-State Hydrogen Deuterium Exchange with Mass Spectrometric Analysis (ssHDX-MS). Pharm Res 35, 12 (2018). https://doi.org/10.1007/s11095-017-2318-9
- monoclonal antibodies
- spray drying