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

## TL;DR

This paper introduces a simulation method to study antibody aggregation and how excipients can inhibit it, reducing the need for costly experiments.

## Contribution

A novel position-restrained simulation method is proposed to study antibody aggregation mechanisms and excipient effects efficiently.

## Key findings

- The simulation method accurately predicts excipient effects on antibody aggregation.
- Results from simulations align with experimental observations on monomer residual content.
- The proposed formulation was validated through experimental characterization.

## Abstract

Background/Objectives: 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. Methods: 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. Results: 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. Conclusions: Simulations and experiments revealed the mechanism and showed consistent trends, providing better understanding for aggregation research.