Temperature and excipient mediated modulation of monoclonal antibody interactions revealed by kD, rheology, and Raman spectroscopy
High-concentration monoclonal antibody (mAb) formulations are frequently constrained by elevated viscosity and colloidal instability, stemming from enhanced intermolecular interactions under crowded conditions. In this study, we delineate the thermodynamic and rheological consequences of modulating...
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Taylor & Francis Group
2025-12-01
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| Series: | mAbs |
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| Online Access: | https://www.tandfonline.com/doi/10.1080/19420862.2025.2521511 |
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| author | Olivia Noelle Eskens Sabitoj Singh Virk Akashdeep Singh Virk Deepika Venkataramani Maureen Crames Maria Calderon Vaca Matthew Matusewicz Connor Smith Joshin George Collin Taylor Michael S. Marlow Samiul Amin |
| author_facet | Olivia Noelle Eskens Sabitoj Singh Virk Akashdeep Singh Virk Deepika Venkataramani Maureen Crames Maria Calderon Vaca Matthew Matusewicz Connor Smith Joshin George Collin Taylor Michael S. Marlow Samiul Amin |
| author_sort | Olivia Noelle Eskens |
| collection | DOAJ |
| description | High-concentration monoclonal antibody (mAb) formulations are frequently constrained by elevated viscosity and colloidal instability, stemming from enhanced intermolecular interactions under crowded conditions. In this study, we delineate the thermodynamic and rheological consequences of modulating protein-protein interactions through excipient-mediated and temperature-dependent mechanisms. Using an orthogonal analytical framework comprising diffusion interaction parameter (kD) measurements, high-shear rheometry, and Raman spectroscopic profiling, we interrogated mAb solutions at ~ 80 and 160 mg/mL across a physiologically and industrially relevant thermal window (5–45 °C). In the absence of ionic additives, high kD values (~60 mL/g) indicated dominant long-range electrostatic repulsions, resulting in suppressed self-association and lower viscosity. Incorporation of NaCl (0.05% w/v) markedly decreased kD (~16–20 mL/g), consistent with Debye screening of surface charges and a shift toward short-range hydrophobic and van der Waals attractions, which became especially pronounced at elevated protein concentrations and lower temperatures. Polysorbate 20 (0.05% v/v) mitigated these interactions via preferential surface adsorption, while sucrose exhibited a dualistic, concentration-dependent influence on viscosity via preferential exclusion and entropic crowding. The combination of NaCl and PS20 yielded the most pronounced rheological suppression, reflecting synergistic attenuation of both long-range repulsion and short-range association. Raman spectral analysis of Amide I/III regions confirmed structural invariance under thermal and shear stress, attributing viscosity modulation to colloidal rather than conformational perturbations. Collectively, these data elucidate the multivariate control of interparticle potentials in mAb solutions and provide a predictive basis for engineering subcutaneous formulations that optimize manufacturability, physical stability, and injectability through strategic manipulation of colloidal interaction landscapes. |
| format | Article |
| id | doaj-art-188d5837b11e409395c037db53e2a75e |
| institution | Kabale University |
| issn | 1942-0862 1942-0870 |
| language | English |
| publishDate | 2025-12-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | mAbs |
| spelling | doaj-art-188d5837b11e409395c037db53e2a75e2025-08-20T03:32:36ZengTaylor & Francis GroupmAbs1942-08621942-08702025-12-0117110.1080/19420862.2025.2521511Temperature and excipient mediated modulation of monoclonal antibody interactions revealed by kD, rheology, and Raman spectroscopyOlivia Noelle Eskens0Sabitoj Singh Virk1Akashdeep Singh Virk2Deepika Venkataramani3Maureen Crames4Maria Calderon Vaca5Matthew Matusewicz6Connor Smith7Joshin George8Collin Taylor9Michael S. Marlow10Samiul Amin11Department of Chemical Engineering, Manhattan University, Riverdale, NY, USADepartment of Chemical, Environmental and Materials Engineering, University of Miami, Miami, FL, USADepartment of Chemical, Environmental and Materials Engineering, University of Miami, Miami, FL, USADepartment of Chemical Engineering, Manhattan University, Riverdale, NY, USABiotherapeutics Discovery Department, Boehringer Ingelheim Pharmaceuticals Inc., Ridgefield, CT, USADepartment of Chemical, Environmental and Materials Engineering, University of Miami, Miami, FL, USADepartment of Chemical Engineering, Manhattan University, Riverdale, NY, USADepartment of Chemical Engineering, Manhattan University, Riverdale, NY, USADepartment of Chemical Engineering, Manhattan University, Riverdale, NY, USADepartment of Chemical, Environmental and Materials Engineering, University of Miami, Miami, FL, USABiotherapeutics Discovery Department, Boehringer Ingelheim Pharmaceuticals Inc., Ridgefield, CT, USADepartment of Chemical, Environmental and Materials Engineering, University of Miami, Miami, FL, USAHigh-concentration monoclonal antibody (mAb) formulations are frequently constrained by elevated viscosity and colloidal instability, stemming from enhanced intermolecular interactions under crowded conditions. In this study, we delineate the thermodynamic and rheological consequences of modulating protein-protein interactions through excipient-mediated and temperature-dependent mechanisms. Using an orthogonal analytical framework comprising diffusion interaction parameter (kD) measurements, high-shear rheometry, and Raman spectroscopic profiling, we interrogated mAb solutions at ~ 80 and 160 mg/mL across a physiologically and industrially relevant thermal window (5–45 °C). In the absence of ionic additives, high kD values (~60 mL/g) indicated dominant long-range electrostatic repulsions, resulting in suppressed self-association and lower viscosity. Incorporation of NaCl (0.05% w/v) markedly decreased kD (~16–20 mL/g), consistent with Debye screening of surface charges and a shift toward short-range hydrophobic and van der Waals attractions, which became especially pronounced at elevated protein concentrations and lower temperatures. Polysorbate 20 (0.05% v/v) mitigated these interactions via preferential surface adsorption, while sucrose exhibited a dualistic, concentration-dependent influence on viscosity via preferential exclusion and entropic crowding. The combination of NaCl and PS20 yielded the most pronounced rheological suppression, reflecting synergistic attenuation of both long-range repulsion and short-range association. Raman spectral analysis of Amide I/III regions confirmed structural invariance under thermal and shear stress, attributing viscosity modulation to colloidal rather than conformational perturbations. Collectively, these data elucidate the multivariate control of interparticle potentials in mAb solutions and provide a predictive basis for engineering subcutaneous formulations that optimize manufacturability, physical stability, and injectability through strategic manipulation of colloidal interaction landscapes.https://www.tandfonline.com/doi/10.1080/19420862.2025.2521511Antibody intermolecular interactionsdiffusion interaction parameterexcipient effectshigh-concentration viscositymonoclonal antibody formulationprotein-protein interactions |
| spellingShingle | Olivia Noelle Eskens Sabitoj Singh Virk Akashdeep Singh Virk Deepika Venkataramani Maureen Crames Maria Calderon Vaca Matthew Matusewicz Connor Smith Joshin George Collin Taylor Michael S. Marlow Samiul Amin Temperature and excipient mediated modulation of monoclonal antibody interactions revealed by kD, rheology, and Raman spectroscopy mAbs Antibody intermolecular interactions diffusion interaction parameter excipient effects high-concentration viscosity monoclonal antibody formulation protein-protein interactions |
| title | Temperature and excipient mediated modulation of monoclonal antibody interactions revealed by kD, rheology, and Raman spectroscopy |
| title_full | Temperature and excipient mediated modulation of monoclonal antibody interactions revealed by kD, rheology, and Raman spectroscopy |
| title_fullStr | Temperature and excipient mediated modulation of monoclonal antibody interactions revealed by kD, rheology, and Raman spectroscopy |
| title_full_unstemmed | Temperature and excipient mediated modulation of monoclonal antibody interactions revealed by kD, rheology, and Raman spectroscopy |
| title_short | Temperature and excipient mediated modulation of monoclonal antibody interactions revealed by kD, rheology, and Raman spectroscopy |
| title_sort | temperature and excipient mediated modulation of monoclonal antibody interactions revealed by kd rheology and raman spectroscopy |
| topic | Antibody intermolecular interactions diffusion interaction parameter excipient effects high-concentration viscosity monoclonal antibody formulation protein-protein interactions |
| url | https://www.tandfonline.com/doi/10.1080/19420862.2025.2521511 |
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