Modulation of the Hydration Water Around Monoclonal Antibodies on Addition of Excipients Detected by Terahertz Time-Domain Spectroscopy.

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Wallace, Vincent P 
Ferachou, Denis 
Ke, Peng 
Day, Katie 
Uddin, Shahid 

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.

analytical biochemistry, arginine, formulation, hydration layer, monoclonal antibody, protein aggregation, protein formulation, stability, terahertz spectroscopy, Antibodies, Monoclonal, Arginine, Chemistry, Pharmaceutical, Excipients, Solutions, Sucrose, Terahertz Spectroscopy, Water
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J Pharm Sci
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Elsevier BV
This work was funded by Innovate UK (grant ref. 131420) and mAbs were supplied by MedImmune. We thank Alessia Portieri and Paddy O’Kelly of Teraview Limited for access to the TPS Spectra 3000 and technical support. The authors confirm that no conflicts of interest exist.