Study of monoclonal antibody aggregation at the air-liquid interface under flow by ATR-FTIR spectroscopic imaging
Author(s)
van Haaren, Céline
Byrne, Bernadette
Kazarian, Sergei G
Type
Journal Article
Abstract
Throughout bioprocessing, transportation, and storage, therapeutic monoclonal antibodies (mAbs) experience stress conditions that may cause protein unfolding and/or chemical modifications. Such structural changes may lead to the formation of aggregates, which reduce mAb potency and may cause harmful immunogenic responses in patients. Therefore, aggregates need to be detected and removed or ideally prevented from forming. Air-liquid interfaces, which arise during various stages of bioprocessing, are one of the stress factors causing mAb aggregation. In this study, the behavior of an immunoglobulin G (IgG) at the air-liquid interface was investigated under flow using macro attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopic imaging. This chemically specific imaging technique allows observation of adsorption of IgG to the air-liquid interface and detection of associated secondary structural changes. Chemical images revealed that IgG rapidly accumulated around an injected air bubble under flow at 45 °C; however, no such increase was observed at 25 °C. Analysis of the second derivative spectra of IgG at the air-liquid interface revealed changes in the protein secondary structure associated with increased intermolecular β-sheet content, indicative of aggregated IgG. The addition of 0.01% w/v polysorbate 80 (PS80) reduced the amount of IgG at the air-liquid interface in a static setup at 30 °C; however, this protective effect was lost at 45 °C. These results suggest that the presence of air-liquid interfaces under flow may be detrimental to mAb stability at elevated temperatures and demonstrate the power of ATR-FTIR spectroscopic imaging for studying the structural integrity of mAbs under bioprocessing conditions.
Date Issued
2024-03-19
Date Acceptance
2024-02-26
Citation
Langmuir: the ACS journal of surfaces and colloids, 2024, 40 (11), pp.5858-5868
ISSN
0743-7463
Publisher
American Chemical Society
Start Page
5858
End Page
5868
Journal / Book Title
Langmuir: the ACS journal of surfaces and colloids
Volume
40
Issue
11
Copyright Statement
Copyright © 2024 The Authors. Published by American Chemical Society. This publication is licensed under
CC-BY 4.0.
CC-BY 4.0.
License URL
Identifier
https://www.ncbi.nlm.nih.gov/pubmed/38445553
Subjects
Antibodies, Monoclonal
Ataxia Telangiectasia Mutated Proteins
Humans
Immunoglobulin G
Protein Structure, Secondary
Protein Unfolding
Spectroscopy, Fourier Transform Infrared
Publication Status
Published
Coverage Spatial
United States
Date Publish Online
2024-03-06