Early detection of protein aggregation in biopharmaceuticals e.g. monoclonal antibodies (mAbs)
Typically monoclonal antibody formulations are made up of monomers. However, during production, filtration and
purification steps, the formulation may start to aggregate and form dimers, trimers, fibrils and/or irregular aspherical
aggregates. This aggregation has negative consequences for the potency, quality and safety of the end product.
Challenge
Biopharmaceutical companies are interested in tools to help monitor aggregation and detect the onset of aggregation levels as soon as possible so the process can be controlled or even stopped when aggregates begin to form.
Experiment
BRAVE Analytics’ nanoparticle analyzer and online particle
sensor provide a way to determine when aggregation
starts to occur. It monitors the progression of aggregation
as it happens via OF2i® and static light scattering signals[1]
and delivers results in real-time, eliminating the delay
inherent in waiting for offline measurements.
[1] Imaging the scattered light of a nanoparticle through a cylindrical capillary. Ulrich Hohenester, Christian Neuper,
Marko Šimić, and Christian Hill. Nanophotonics, 13(4):457–463, 2024.
Fig. 1: OF2i® single-particle light scattering model: OF2i®-based calculation of aggregation states to determine aggregation thresholds
Fig. 2: Comparison of protein aggregation: static light scattering intensity mean and object count over time
Benefits of OF2i®
The BRAVE B-Aware module of the BRAVE B-Curious nanoparticle analyzer determines scattering from a very defined scanning volume (Fig 1.). This scattering signal is used to calculate aggregation thresholds and determine larger objects (e.g. fibrils) on a single-particle-based resolution (counting-based method). At known concentrations, OF2i® physical models are able to give absolute aggregation values and/or provide critical quality attributes (CQA) continously and in real time.
Configuration of the setup:
BRAVE B-Curious device with BRAVE B-Aware software module for particle sizing, Fluid Automation Module, Control and Evaluation Module and software
The analysis setup used in this case
The BRAVE B-Curious particle analyzer with BRAVE B-Aware software module.
The BRAVE B-Aware module detects and tracks very small particles (from 5 nm) and delivers particle counts and particle concentration values.
Ideal for detecting the onset of aggregation and tracking sample behavior when very small particles are involved.
Continuous in-flow measurement.
