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In-Solution IgG Titer Determination in Fermentation Broth Using Affibodies and Flow-Induced Dispersion Analysis
Morten E Pedersen1,2, Jesper Østergaard2, Henrik Jensen1,2
1FIDA Biosystems ApS, Fruebjesrgvej 3, 2100 Copenhagen, Denmark.
ACS Omega
|May 20, 2020
Summary
Flow-induced dispersion analysis (FIDA) enables rapid, in-solution quantification of monoclonal IgG antibodies like rituximab in cell culture. This novel method offers a faster alternative to traditional assays for biopharmaceutical process monitoring.
Area of Science:
- Biopharmaceutical analysis
- Analytical chemistry
- Biotechnology
Background:
- Biopharmaceutical production requires monitoring of protein and peptide-based drugs during fermentation and for release testing.
- Current methods like ELISA and chromatography are multi-step, lengthy, and impractical for real-time process control.
- Novel in-solution techniques are needed for faster, more efficient biopharmaceutical analysis.
Purpose of the Study:
- To demonstrate the utility of flow-induced dispersion analysis (FIDA) for quantifying monoclonal IgG antibodies directly in cell fermentation broth.
- To establish FIDA as a rapid and efficient method for biopharmaceutical process monitoring.
- To characterize the binding kinetics and complex formation using FIDA.
Main Methods:
- Utilized flow-induced dispersion analysis (FIDA), an in-solution ligand binding technology based on Taylor dispersion analysis.
- Employed a fluorophore-conjugated anti-IgG affibody as a selective binder for rituximab.
- Generated a binding curve by measuring the change in hydrodynamic radius of the affibody upon interaction with known rituximab concentrations in fermentation medium.
Main Results:
- Successfully quantified monoclonal IgG antibodies (rituximab) in mammalian cell fermentation broth with an analysis time of only 4 minutes.
- Determined the dissociation constant and complex size of the rituximab-affibody interaction.
- Demonstrated the feasibility of using FIDA for real-time monitoring and titer determination in bioprocesses.
Conclusions:
- Flow-induced dispersion analysis (FIDA) provides a rapid, sensitive, and efficient method for quantifying monoclonal IgG antibodies in complex biological matrices like fermentation broth.
- FIDA overcomes limitations of traditional surface-based assays, enabling practical real-time process control in biopharmaceutical manufacturing.
- This technology offers a significant advancement for the analysis of protein-based therapeutics, reducing analysis time and sample volume requirements.
