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Pharmaceutical Research

, 35:12 | Cite as

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

  • Ehab M. Moussa
  • Nathan E. Wilson
  • Qi Tony Zhou
  • Satish K. Singh
  • Sandeep Nema
  • Elizabeth M. ToppEmail author
Research Paper Theme: Formulation and Manufacturing of Solid Dosage Forms
Part of the following topical collections:
  1. Formulation and Manufacturing of Solid Dosage Forms

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.

KEY WORDS

conformation interactions lyophilization monoclonal antibodies spray drying 

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

Notes

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.

Supplementary material

11095_2017_2318_MOESM1_ESM.docx (1005 kb)
ESM 1 (DOCX 1005 kb)

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Ehab M. Moussa
    • 1
  • Nathan E. Wilson
    • 1
  • Qi Tony Zhou
    • 1
  • Satish K. Singh
    • 2
    • 3
  • Sandeep Nema
    • 2
  • Elizabeth M. Topp
    • 1
    Email author
  1. 1.Department of Industrial and Physical Pharmacy, College of PharmacyPurdue UniversityWest LafayetteUSA
  2. 2.BioTherapeutics Pharmaceutical SciencesPfizer Inc.ChesterfieldUSA
  3. 3.Lonza AGBaselSwitzerland

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