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Biophysical and Sequence-Based Methods for Identifying Monovalent and Bivalent Antibodies with High Colloidal
Magfur E Alam1, Steven B Geng1, Christian Bender2
1Isermann Department of Chemical & Biological Engineering, Center for Biotechnology & Interdisciplinary Studies, Rensselaer Polytechnic Institute , Troy, New York 12180, United States.
Molecular Pharmaceutics
|November 21, 2017
Summary
We developed a new method, self-interaction nanoparticle spectroscopy (SINS), to predict antibody self-association early in discovery. This helps identify antibody fragments (Fabs) that will have good properties when reformatted as monoclonal antibodies (mAbs).
Area of Science:
- Biochemistry
- Biotechnology
- Immunology
Background:
- Antibody fragments (Fabs) are used for discovery but reformatted into bivalent monoclonal antibodies (mAbs) for therapy.
- Bivalency in mAbs can cause self-association, leading to poor biophysical properties like low solubility and high viscosity.
- Early identification of Fabs with favorable properties upon reformatting is crucial for therapeutic antibody development.
Purpose of the Study:
- To develop a facile method for evaluating Fab self-association in a multivalent format.
- To identify Fabs that exhibit low self-association and favorable colloidal properties when reformatted as bivalent mAbs.
- To correlate Fab self-association with bivalent mAb self-interaction and biophysical properties.
Main Methods:
- Developed self-interaction nanoparticle spectroscopy (SINS) to assess Fab self-association.
- Immobilized Fabs on gold nanoparticles in a multivalent format.
- Evaluated self-association via plasmon wavelength shifts and absorbance changes.
Main Results:
- SINS measurements of Fab self-association correlated with self-interaction measurements of bivalent mAbs.
- Identified antibodies with favorable biophysical properties using SINS.
- Physicochemical properties of complementarity-determining regions (CDRs), particularly net charge, significantly influence antibody self-association.
Conclusions:
- SINS is a useful tool for predicting mAb biophysical properties from Fabs.
- CDR properties, especially positive charge, are key determinants of antibody self-association.
- Findings aid in developing therapeutic antibodies suitable for high-concentration applications.
