Both Reversible Self-Association and Structural Changes Underpin Molecular Viscoelasticity of mAb Solutions
| Autor: | Gary G. Adams, Stephen E. Harding, Prakash Manikwar, Tabot M. D. Besong, Maria Monica Castellanos, Prasad Sarangapani, Justin Weaver, Jai A. Pathak, Steven M. Bishop, Arun Parupudi |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Circular dichroism Hydrodynamic radius Size-exclusion chromatography Analytical chemistry Pharmaceutical Science CHO Cells 030226 pharmacology & pharmacy Viscoelasticity 03 medical and health sciences Viscosity Cricetulus 0302 clinical medicine Differential scanning calorimetry Rheology Dynamic light scattering Cricetinae Animals Protein Interaction Domains and Motifs Calorimetry Differential Scanning Chemistry Antibodies Monoclonal Elasticity Pharmaceutical Solutions 030104 developmental biology Chromatography Gel |
| Zdroj: | Journal of Pharmaceutical Sciences. 105:3496-3506 |
| ISSN: | 0022-3549 |
| DOI: | 10.1016/j.xphs.2016.08.020 |
| Popis: | The role of antibody structure (conformation) in solution rheology is probed. It is demonstrated here that pH-dependent changes in the tertiary structure of 2 mAb solutions lead to viscoelasticity and not merely a shear viscosity (η) increase. Steady shear flow curves on mAb solutions are reported over broad pH (3.0 ≤ pH ≤ 8.7) and concentration (2 mg/mL ≤ c ≤ 120 mg/mL) ranges to comprehensively characterize their rheology. Results are interpreted using size exclusion chromatography, differential scanning calorimetry, analytical ultracentrifugation, near-UV circular dichroism, and dynamic light scattering. Changes in tertiary structure with concentration lead to elastic yield stress and increased solution viscosity in solution of “mAb1.” These findings are supported by dynamic light scattering and differential scanning calorimetry, which show increased hydrodynamic radius of mAb1 at low pH and a reduced melting temperature Tm, respectively. Conversely, another molecule at 120 mg/mL solution concentration is a strong viscoelastic gel due to perturbed tertiary structure (seen in circular dichroism) at pH 3.0, but the same molecule responds as a viscous liquid due to reversible self-association at pH 7.4 (verified by analytical ultracentrifugation). Both protein–protein interactions and structural perturbations govern pH-dependent viscoelasticity of mAb solutions. |
| Databáze: | OpenAIRE |
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