Aggregation in drug development: a challenge that can be avoided

Aggregation in drug development

In rare situations, aggregation may be intentionally utilised as a therapeutic strategy, but in most cases, the objective is to prevent or minimise aggregation due to its potential negative effects on drug stability, efficacy, and safety. Thus, the formation of aggregates often presents a significant challenge in drug development, including drug formulation and bioprocessing.

In this article from the FIDA Quality Control Series, we will highlight how protein aggregation can be quantitatively measured to indicate compromised proteins and unravel the root causes of aggregation.

Why does protein aggregation occur?

In drug development, aggregation can be caused by various factors. It can be due to the inherent physicochemical properties of the molecules, such as hydrophobic and hydrophilic interactions or conformational instability, including misfolding. It can also be caused by formulation-related factors, including high drug concentrations, or by external conditions such as changes in pH or temperature, or environmental changes such as ionic strength or additives. Furthermore, it can be induced by mechanical stress, e.g., filtration or stirring, improper storage, or the presence of impurities. The ability to detect and mitigate aggregation throughout the life cycle of the molecule is of high importance to both academia and industry, thus the question is...

How can protein aggregation be measured?

Standard biochemical methods for identifying protein aggregates, such as centrifugation, size-exclusion chromatography, and gel electrophoresis, are not ideal for in-depth management of aggregation. They are not universally applicable to all samples and often require optimisation. None can accurately identify, count, and quantify the size of aggregates. Many of the current methods also require purifying the protein under certain pH, temperature, and/or buffer conditions, which prevents the study of how these factors may impact aggregation. A high-throughput, quantitative, and in-solution method is needed to screen for aggregates, identify the source of aggregation, and develop strategies to prevent aggregation in both human diseases and protein production.

A high-throughput, quantitative, and in-solution method is needed to screen for aggregates, identify the source of aggregation, and develop strategies to prevent aggregation in both human diseases and protein production.

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Why are biophysical methods a good choice for measurement of protein aggregation?

Biophysical methods provide a transparent, high-throughput, and quantitative measurement of protein aggregation compared to biochemical approaches.

FIDA employs a biophysical method to measure insoluble aggregates in a solution. When the sample is flowed through a FIDA capillary, aggregates are visible as distinct spikes in the raw FIDA signal. The FIDA software readout will provide information on the number of aggregates within a sample. And there is no need to purify protein from the matrix, no matter its complexity, providing valuable information on the role of the buffer in aggregation. Additionally, FIDA’s built-in autosampler can autonomously screen buffers to see how they impact aggregation, providing a valuable, timesaving tool for protein assays. Would you like to know more? Do not hesitate to ask a question to one of our scientists.

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If you are studying biomolecular condensates we recommend you to read more here.

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