Viscosities and Protein Interactions of Bispecific Antibodies and Their Monospecific Mixtures
| Autor: | Danielle L. Leiske, Eric M. Furst, Christopher J. Roberts, William J. Galush, Mahlet A. Woldeyes, Lilian Lam Josephson |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Light medicine.drug_class Diffusion Pharmaceutical Science Ionic bonding 02 engineering and technology Monoclonal antibody 03 medical and health sciences Viscosity symbols.namesake Dynamic light scattering Antibodies Bispecific Drug Discovery medicine Molecule Arrhenius equation Chemistry Osmolar Concentration Antibodies Monoclonal Proteins 021001 nanoscience & nanotechnology Crystallography 030104 developmental biology Isoelectric point symbols Molecular Medicine 0210 nano-technology Protein Binding |
| Zdroj: | Molecular Pharmaceutics. 15:4745-4755 |
| ISSN: | 1543-8392 1543-8384 |
| Popis: | Solution viscosities (η) and protein-protein interactions (PPI) of three monoclonal antibodies (mAb-A, mAb-B, mAb-C), two bispecific antibodies (BsAb-A/B, BsAb-A/C), and two 1:1 binary mixtures (mAb-A + mAb-B and mAb-A + mAb-C) were measured. mAb-A and mAb-C have similar isoelectric point (pI) values but significantly different η versus protein concentration ( c2) profiles. The viscosity of the mAb-A + mAb-C mixture followed an Arrhenius mixing rule and was identical to viscosity of the bispecific BsAb-A/C. In contrast, mAb-A and mAb-B had similar η versus c2 profiles, but the Arrhenius mixing rule failed to predict the higher viscosities of their mixtures. The viscosity of the bispecific BsAb-A/B followed the 1:1 mAb-A + mAb-B mixture at all concentrations. The nature of the interactions for mAb-A, mAb-B, the BsAb-A/B bispecific, and the 1:1 mAb-A + mAb-B mixture were characterized by static and dynamic light scattering (SLS and DLS). mAb-A and mAb-B exhibited net-attractive and -repulsive electrostatic interactions, respectively. The bispecific antibody (BsAb-A/B) had short-ranged attractive interactions, suggesting that the increase in viscosity for this molecule and the mAb-A + mAb-B mixture was due to cross-interactions between Fab regions. At high and low ionic strengths and protein concentrations, the Rayleigh scattering profile, the collective diffusion coefficient, and viscosity for the mixture closely followed that for the bispecific antibody. These results highlight the possible anomalous viscosity increases of bispecific antibodies constructed from relatively low-viscosity mAbs but demonstrates a potentially fruitful approach of using mAb mixtures to predict the viscosity of candidate bispecific constructs. |
| Databáze: | OpenAIRE |
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