Modulation of the Hydration Water Around Monoclonal Antibodies on Addition of Excipients Detected by Terahertz Time-Domain Spectroscopy.
Terahertz time-domain spectroscopy (THz-TDS) has been shown to detect overlapping extended hydration layers around proteins. Here, we used THz-TDS to detect modulation of the extended hydration layer around monoclonal antibodies (mAbs) by the introduction of commonly used excipients. Proline and sucrose altered the hydration layer around a mAb (mAb1), which was observed as a negative shift in the plateau in absorbance above ~100 mg/mL mAb1 (~70,000 water molecules per mAb); arginine had no effect. At lower concentrations of ~10 mg/mL mAb1 (~700,000 water molecules per mAb) proline and arginine modulated the hydration layer, which was observed as a negative shift in the relative absorbance, whereas sucrose had no effect. The changes in the extended hydration layer were not translated to shifts in the thermal stability or protein:protein interaction parameter. The hydration layer of a second mAb (mAb2) was further shown to be modulated by more complex formulations composed of two or more excipients; although the differences in terahertz absorbance were not predictive of viscosity or long-term stability. THz-TDS promises to be a useful tool for understanding a protein's interaction with excipients in solution and the challenge will be to determine how to apply this knowledge to protein formulation.
School of Physics, University of Western Australia, Crawley, Western Australia 6009, Australia.
Vincent P
VP
Wallace
Department of Chemical Engineering & Biotechnology, University of Cambridge, Cambridge CB2 3RA, UK.
Denis
D
Ferachou
Formulation Sciences, MedImmune Ltd., Cambridge CB21 6GH, UK.
Peng
P
Ke
Formulation Sciences, MedImmune Ltd., Cambridge CB21 6GH, UK.
Katie
K
Day
Formulation Sciences, MedImmune Ltd., Cambridge CB21 6GH, UK.
Shahid
S
Uddin
Analytical Biochemistry, MedImmune LLC, Gaithersburg, Maryland 20878.
Jose
J
Casas-Finet
Formulation Sciences, MedImmune Ltd., Cambridge CB21 6GH, UK.
Christopher F
CF
Van Der Walle
Department of Chemical & Biological Engineering, ChELSI Institute, University of Sheffield, Sheffield S1 3JD, UK. Electronic address: r.j.falconer@sheffield.ac.uk.
Robert J
RJ
Falconer
Department of Chemical Engineering & Biotechnology, University of Cambridge, Cambridge CB2 3RA, UK.
J Axel
JA
Zeitler
D000911:Antibodies, Monoclonal / Q000737:chemistry*; D001120:Arginine / Q000737:chemistry; D002626:Chemistry, Pharmaceutical / Q000379:methods; D005079:Excipients / Q000737:chemistry*; D012996:Solutions / Q000737:chemistry; D013395:Sucrose / Q000737:chemistry; D055068:Terahertz Spectroscopy / Q000379:methods; D014867:Water / Q000737:chemistry*
D000911:Antibodies, Monoclonal; D005079:Excipients; D012996:Solutions; D014867:Water; D013395:Sucrose; D001120:Arginine
analytical biochemistry; arginine; formulation; hydration layer; monoclonal antibody; protein aggregation; protein formulation; stability; terahertz spectroscopy