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

Area of Science:

• Biopharmaceutical Manufacturing
• Analytical Chemistry
• Materials Science

Background:

• Monitoring biopharmaceutical critical quality attributes is essential for Process Analytical Technology (PAT) and Quality by Design (QbD).
• Controlling protein aggregation is vital to prevent immunogenic responses and ensure drug efficacy.
• Existing methods for protein aggregation characterization can be time-consuming or lack sensitivity.

Purpose of the Study:

• To develop and validate hydrogel-encapsulated, label-free fluorescent nanosensors for characterizing protein aggregation.
• To utilize mathematical modeling to understand sensor response and optimize hydrogel properties.
• To demonstrate the capability of these nanosensors for sensitive detection of protein aggregates.

Main Methods:

• Development of hydrogel-encapsulated fluorescent nanosensors.
• Application of mathematical modeling to describe sensor-analyte interactions and hydrogel performance.
• Testing with UV-stressed and light-stressed IgG1 samples.
• Correlation of nanosensor data with Size-Exclusion Chromatography (SEC) results.

Main Results:

• The nanosensors demonstrated label-free detection of protein aggregation.
• Mathematical modeling accurately described the dynamic response of the sensors.
• The system successfully characterized UV-stressed and light-stressed IgG1 samples.
• Detection of high-molecular-weight protein species at concentrations as low as 1% was achieved.

Conclusions:

• Hydrogel-encapsulated fluorescent nanosensors offer a promising tool for biopharmaceutical characterization.
• This technology represents a significant advancement toward rapid Process Analytical Technologies (PAT).
• The developed method facilitates sensitive and label-free monitoring of protein aggregation, crucial for ensuring drug quality and safety.