Nucleation in Protein Aggregation in Biotherapeutic Development: A look into the Heart of the Event

Das, Tapan K.; Chou, Danny K.; Jiskoot, Wim; Arosio, Paolo

doi:10.1016/j.xphs.2022.01.017

Cited by 11 publications

(7 citation statements)

References 135 publications

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“…Often aggregation is triggered by a chemical or conformational change of the antibody structure and is therefore defined as non-native. 100 …”

Section: Orthogonal Approaches For the Selection Of Drug-like Antibodiesmentioning

confidence: 99%

“…This is partly due to the complex nature of this degradation pathway that includes multiple elementary steps of nucleation and aggregate growth that are dependent on both structural (e.g., partial unfolding) and colloidal aspects. 100 The aggregation mechanisms substantially vary between antibodies and are strongly dependent on extrinsic factors such as temperature and pH. 31 , 101 , 102 As a consequence, accelerated stability studies at 40°C or higher temperatures may not always be representative of the stability of antibodies at lower storage temperatures.…”

Section: Orthogonal Approaches For the Selection Of Drug-like Antibodiesmentioning

confidence: 99%

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Approaches to expand the conventional toolbox for discovery and selection of antibodies with drug-like physicochemical properties

Svilenov

Arosio

Menzen³

et al. 2023

mAbs

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Antibody drugs should exhibit not only high-binding affinity for their target antigens but also favorable physicochemical drug-like properties. Such drug-like biophysical properties are essential for the successful development of antibody drug products. The traditional approaches used in antibody drug development require significant experimentation to produce, optimize, and characterize many candidates. Therefore, it is attractive to integrate new methods that can optimize the process of selecting antibodies with both desired target-binding and drug-like biophysical properties. Here, we summarize a selection of techniques that can complement the conventional toolbox used to de-risk antibody drug development. These techniques can be integrated at different stages of the antibody development process to reduce the frequency of physicochemical liabilities in antibody libraries during initial discovery and to co-optimize multiple antibody features during early-stage antibody engineering and affinity maturation. Moreover, we highlight biophysical and computational approaches that can be used to predict physical degradation pathways relevant for long-term storage and in-use stability to reduce the need for extensive experimentation.

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“…Often aggregation is triggered by a chemical or conformational change of the antibody structure and is therefore defined as non-native. 100 …”

Section: Orthogonal Approaches For the Selection Of Drug-like Antibodiesmentioning

confidence: 99%

Section: Orthogonal Approaches For the Selection Of Drug-like Antibodiesmentioning

confidence: 99%

Approaches to expand the conventional toolbox for discovery and selection of antibodies with drug-like physicochemical properties

Svilenov

Arosio

Menzen³

et al. 2023

mAbs

Self Cite

View full text Add to dashboard Cite

show abstract

“…As a result, much effort has been dedicated to understanding the mechanisms of non-native protein aggregation. Currently, bulk-mediated aggregation pathways are well understood, and it is known that conformational stability impacts protein aggregation. − Chemical stresses such as pH and ionic strength, and mechanical stresses such as temperature, multiple freeze–thaw cycles, and the application of shear are all known to initiate aggregation. More recently, it has been established that the presence of interfaces constitutes an additional source of stress during biotherapeutics manufacturing, shelf life, and administration. , Air–liquid, solid–liquid, and liquid–liquid interfaces can be found during chromatography purification, filtration, pumping, and finishing steps.…”

Section: Introductionmentioning

confidence: 99%

Correlating Surface Activity with Interface-Induced Aggregation in a High-Concentration mAb Solution

Escobar,

Griffin,

Dhar

2024

Mol. Pharmaceutics

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Interface-induced aggregation resulting in protein particle formation is an issue during the manufacturing and storage of protein-based therapeutics. High-concentration formulations of therapeutic proteins are even more prone to protein particle formation due to increased protein−protein interactions. However, the dependence of interface-induced protein particle formation on bulk protein concentration is not understood. Furthermore, the formation of protein particles is often mitigated by the addition of polysorbate-based surfactants. However, the details of surfactantprotein interactions that prevent protein particle formation at high concentrations remain unclear. In this work, a tensiometer technique was used to evaluate the surface pressure of an industrially relevant mAb at different bulk concentrations, and in the absence and presence of a polysorbate-based surfactant, polysorbate 20 (PS20). The adsorption kinetics was correlated with subvisible protein particle formation at the air−water interface and in the bulk protein solution using a microflow imaging technique. Our results showed that, in the absence of any surfactant, the number of subvisible particles in the bulk protein solutions increased linearly with mAb concentration, while the number of protein particles measured at the interface showed a logarithmic dependence on bulk protein concentration. In the presence of surfactants above the critical micelle concentration (CMC), our results for lowconcentration mAb solutions (10 mg/mL) showed an interface that is surfactant-dominated, and particle characterization results showed that the addition of the surfactant led to reduced particle formation. In contrast, for the highest concentration (170 mg/mL), coadsorption of proteins and surfactants was observed at the air−water interface, even for surfactant formulations above CMC and the surfactant did not mitigate subvisible particle formation. Our results taken together provide evidence that the ratio between the surfactant and mAb molecules is an important consideration when formulating high-concentration mAb therapeutics to prevent unwanted aggregation

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“…The mechanisms at play differ from protein to protein and the conditions to which the protein is exposed. In addition, multiple pathways may occur simultaneously in any given system. , Typically, protein aggregation involves some degree of conformational change at the monomer or oligomer level, leading to irreversible formation of non-native structures …”

Section: Introductionmentioning

confidence: 99%

Study of Monoclonal Antibody Aggregation at the Air–Liquid Interface under Flow by ATR-FTIR Spectroscopic Imaging

van Haaren,

Byrne,

Kazarian

2024

Langmuir

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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.

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Nucleation in Protein Aggregation in Biotherapeutic Development: A look into the Heart of the Event

Cited by 11 publications

References 135 publications

Approaches to expand the conventional toolbox for discovery and selection of antibodies with drug-like physicochemical properties

Approaches to expand the conventional toolbox for discovery and selection of antibodies with drug-like physicochemical properties

Correlating Surface Activity with Interface-Induced Aggregation in a High-Concentration mAb Solution

Study of Monoclonal Antibody Aggregation at the Air–Liquid Interface under Flow by ATR-FTIR Spectroscopic Imaging

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