Measurements

Aggregation

Protein aggregation results from irreversible conformational changes and misfolding at the structural level which remains a concern and challenge to biotherapeutic development. Typical indicators of aggregation include shifting ratios of alpha-helix and beta-sheet moieties with an increase in anti-parallel beta-sheet structures typically corresponding to an increase in intermolecular protein aggregation. This occurrence directly relates to potential detrimental effects on stability, efficacy, and safety due to the precipitation of insoluble populations. Thus, it remains important to detect and prevent protein aggregation to protect the integrity of a biotherapeutic molecule within the development and formulation processes.  

By incorporating the novel secondary structural characterization tool Microfluidic Modulation Spectroscopy (MMS) into the biomolecular characterization workflow, it is possible to detect and track significant markers of aggregation and instability. This is accomplished by following changes in both parallel and anti-parallel beta-sheet content across an entire formulation experiment to pinpoint the conditions where aggregation started and increased. The ability to inform upstream development decisions with early and actionable results is invaluable, and with MMS, stability monitoring is easy and purpose-built into the platform to protect downstream processing.  

Higher Order Structure (HOS) populations measured in a pressure treated aggregated IgG1 spike series analyzed by MMS. As the concentration of aggregated IgG1 was increased from 0% to 100%, the respective and expected increases in anti-parallel beta-sheet (beta-) and decreases in parallel beta-sheet (beta) were accurately measured. Application Note Detection of Pressure Induced Aggregation using MMS.

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