Characterization of Protein Aggregation Using Hydrogel-Encapsulated nIR Fluorescent Nanoparticle Sensors.

The monitoring of biopharmaceutical critical quality attributes in-process, at both the process development and manufacturing stages, is necessary for the implementation of process analytical technology and quality-by-design principles. Among these attributes, it is important to monitor and control protein aggregation during the manufacturing of biological therapeutics to prevent adverse immunogenic responses and minimize negative impacts on drug deliverability. In this work, we explore hydrogel-encapsulated, label-free fluorescent nanosensors for the characterization of protein aggregation. A mathematical model is used to describe the diffusion and binding of a series of stressed pharmaceutical samples to such sensors, describing their dynamic response. We use mathematical modeling to map the influence of hydrogel properties on the separation performance, given the composition of UV-stressed IgG1 samples. Using this modified model, the compositions of light-stressed IgG1 samples were fit to experimental data and correlated with size-exclusion chromatography data. The results demonstrate the ability to detect the presence of high-molecular-weight protein species at a concentration as low as 1%. This work represents a significant step toward the development and deployment of rapid process analytical technologies for biopharmaceutical characterization.