A Convenient Strategy for Studying Antibody Aggregation and Inhibition of Aggregation: Characterization and Simulation

<b>Background/Objectives:</b> Protein aggregation, particularly the aggregation of antibody-based drugs, has long been a significant challenge in downstream processes and formulation. While the inhibitory effects of excipients on aggregation have been extensively studied using early expe...

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Main Authors: Yibo Guo, Xi Chen, Guchen Fang, Xuejun Cao, Junfen Wan
Format: Article
Language:English
Published: MDPI AG 2025-04-01
Series:Pharmaceutics
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Online Access:https://www.mdpi.com/1999-4923/17/4/534
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author Yibo Guo
Xi Chen
Guchen Fang
Xuejun Cao
Junfen Wan
author_facet Yibo Guo
Xi Chen
Guchen Fang
Xuejun Cao
Junfen Wan
author_sort Yibo Guo
collection DOAJ
description <b>Background/Objectives:</b> Protein aggregation, particularly the aggregation of antibody-based drugs, has long been a significant challenge in downstream processes and formulation. While the inhibitory effects of excipients on aggregation have been extensively studied using early experimental characterization methods, complete formulation research requires significant amounts of antibodies and time, resulting in high research costs. <b>Methods:</b> This study proposed a quick and small-scale position-restrained simulation method which elucidated the mechanism of the reversible self-association (RSA) of antibodies and the influence of excipients on RSA under different conditions. We also validated the rationality of rapid and small-scale simulations through long-term (>1 μs) and large-scale (>1,000,000 atoms) simulations. <b>Results</b>: Through combing with simple stability characterization, the effects of different excipients on monomer residual content and the trend shown with concentration changes after thermal incubation were found to be similar to those observed in the simulations. Additionally, the formulation proposed by the simulations was validated using experimental characterization. <b>Conclusions</b>: Simulations and experiments revealed the mechanism and showed consistent trends, providing better understanding for aggregation research.
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spelling doaj-art-d59e09abf8094f1aa94769c626e189bd2025-08-20T03:13:51ZengMDPI AGPharmaceutics1999-49232025-04-0117453410.3390/pharmaceutics17040534A Convenient Strategy for Studying Antibody Aggregation and Inhibition of Aggregation: Characterization and SimulationYibo Guo0Xi Chen1Guchen Fang2Xuejun Cao3Junfen Wan4State Key Laboratory of Bioreactor Engineering, Department of Bioengineering, East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, ChinaState Key Laboratory of Bioreactor Engineering, Department of Bioengineering, East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, ChinaState Key Laboratory of Bioreactor Engineering, Department of Bioengineering, East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, ChinaState Key Laboratory of Bioreactor Engineering, Department of Bioengineering, East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, ChinaState Key Laboratory of Bioreactor Engineering, Department of Bioengineering, East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, China<b>Background/Objectives:</b> Protein aggregation, particularly the aggregation of antibody-based drugs, has long been a significant challenge in downstream processes and formulation. While the inhibitory effects of excipients on aggregation have been extensively studied using early experimental characterization methods, complete formulation research requires significant amounts of antibodies and time, resulting in high research costs. <b>Methods:</b> This study proposed a quick and small-scale position-restrained simulation method which elucidated the mechanism of the reversible self-association (RSA) of antibodies and the influence of excipients on RSA under different conditions. We also validated the rationality of rapid and small-scale simulations through long-term (>1 μs) and large-scale (>1,000,000 atoms) simulations. <b>Results</b>: Through combing with simple stability characterization, the effects of different excipients on monomer residual content and the trend shown with concentration changes after thermal incubation were found to be similar to those observed in the simulations. Additionally, the formulation proposed by the simulations was validated using experimental characterization. <b>Conclusions</b>: Simulations and experiments revealed the mechanism and showed consistent trends, providing better understanding for aggregation research.https://www.mdpi.com/1999-4923/17/4/534protein aggregationMD simulationbispecific antibodysingle-chain fragment variable
spellingShingle Yibo Guo
Xi Chen
Guchen Fang
Xuejun Cao
Junfen Wan
A Convenient Strategy for Studying Antibody Aggregation and Inhibition of Aggregation: Characterization and Simulation
Pharmaceutics
protein aggregation
MD simulation
bispecific antibody
single-chain fragment variable
title A Convenient Strategy for Studying Antibody Aggregation and Inhibition of Aggregation: Characterization and Simulation
title_full A Convenient Strategy for Studying Antibody Aggregation and Inhibition of Aggregation: Characterization and Simulation
title_fullStr A Convenient Strategy for Studying Antibody Aggregation and Inhibition of Aggregation: Characterization and Simulation
title_full_unstemmed A Convenient Strategy for Studying Antibody Aggregation and Inhibition of Aggregation: Characterization and Simulation
title_short A Convenient Strategy for Studying Antibody Aggregation and Inhibition of Aggregation: Characterization and Simulation
title_sort convenient strategy for studying antibody aggregation and inhibition of aggregation characterization and simulation
topic protein aggregation
MD simulation
bispecific antibody
single-chain fragment variable
url https://www.mdpi.com/1999-4923/17/4/534
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