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Effects of Drying Process on an IgG1 Monoclonal Antibody Using Solid-State Hydrogen Deuterium Exchange with Mass Spectrometric Analysis (ssHDX-MS)

  • Research Paper
  • Theme: Formulation and Manufacturing of Solid Dosage Forms
  • Published:
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Abstract

Purpose

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).

Methods

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.

Results

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.

Conclusions

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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Abbreviations

ATR:

Attenuated total reflectance

BSA:

Bovine serum albumin

CD:

Circular dichroism

CN:

Controlled ice nucleation

FTIR:

Fourier transform infrared

HDX-MS:

Hydrogen deuterium exchange with mass spectrometric analysis

IgG :

Immunoglobulin G

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

mAb:

Monoclonal antibody

Nfast :

Number of amide hydrogen atoms in the fast exchanging pool

Nslow :

Number of amide hydrogen atoms in the slow exchanging pool

PMT:

Photomultiplier tube

PXRD:

Powder x-ray diffraction

SD:

Spray drying

SEM:

Scanning electron microscope

ss:

Solid-state; indicates that an analytical method has been applied to a solid sample

ssPL-MS :

Solid-state photolytic labeling with mass spectrometric analysis

UNC:

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.

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Author notes

  1. Satish K. Singh

    Present address: Lonza AG, Basel, Switzerland

Authors and Affiliations

  1. Department of Industrial and Physical Pharmacy, College of Pharmacy, Purdue University, 575 Stadium Mall Drive, West Lafayette, Indiana, 47906, USA

    Ehab M. Moussa, Nathan E. Wilson, Qi Tony Zhou & Elizabeth M. Topp

  2. BioTherapeutics Pharmaceutical Sciences, Pfizer Inc., Chesterfield, Missouri, USA

    Satish K. Singh & Sandeep Nema

Authors
  1. Ehab M. Moussa
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  2. Nathan E. Wilson
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  3. Qi Tony Zhou
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  6. Elizabeth M. Topp
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Corresponding author

Correspondence to Elizabeth M. Topp.

Additional information

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

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  • DOI: https://doi.org/10.1007/s11095-017-2318-9

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