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Effects of Histidine and Sucrose on the Biophysical Properties of a Monoclonal Antibody

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Abstract

Purpose

Histidine is a commonly used buffer in formulation of monoclonal antibodies (mAb), often with excipients like sucrose. The objective of this study was to examine the effects of both histidine and sucrose on the biophysical characteristics of a mAb.

Methods

The hydrodynamic radius of the mAb was determined by dynamic light scattering and confirmed by size exclusion chromatography. Data were also obtained for the osmotic virial coefficients (from osmotic pressure measurements), the solution viscosity, and the mAb thermal stability (using differential scanning calorimetry) at selected conditions.

Results

There were no significant changes in mAb conformation / stability as determined by DSC. The hydrodynamic radius initially increased with increasing histidine concentration, going through a maximum at a histidine concentration of about 20 mM. The addition of sucrose increased the mAb hydrodynamic radius at all histidine concentrations by about 0.5 nm. The observed effects of histidine and sucrose on the hydrodynamic radius were also reflected in changes in the osmotic pressure and solution viscosity.

Conclusions

These results provide important insights into the effects of both histidine and sucrose on the behavior of concentrated mAb solutions, including the potential impact on ultrafiltration / diafiltration processes.

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Abbreviations

ε:

Permittivity of medium (C2 s2 / kg / m3)

ζ:

Surface potential (V)

κ:

Inverse Debye length (m−1)

η:

Viscosity of mAb solution (Pa s)

μ:

Solvent viscosity (Pa s)

∆P:

Pressure difference (Pa)

ΔΠ:

Osmotic pressure difference between the solutions (Pa)

σo :

Osmotic reflection coefficient

b:

Viscosity parameter (L/g)

Bi :

Osmotic virial coefficients

Cmax :

Maximum protein concentration in viscosity correlation (g/L)

Cp :

mAb concentration

DLS:

Dynamic light scattering

DSC:

Differential scanning calorimetry

Jv :

Filtrate flux (L/m2 h)

L:

Capillary length (m)

Lp :

Membrane Permeability (L/m2 h bar)

mAb:

Monoclonal Antibody

Mp :

Molecular weight of the mAb (Da)

ms :

Molar salt concentration (mM)

MW:

Dextran molecular weight (Da)

Q:

Volumetric flow rate (m3)

R:

Capillary radius (m)

rs :

Hydrodynamic radius (nm)

SEC:

Size exclusion chromatography

T:

Absolute temperature (K)

Tm :

Melting temperature in differential scanning calorimetry (K)

UF:

Ultrafiltration

Z:

Net protein charge

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ACKNOWLEDGMENTS AND DISCLOSURES

The authors would like to acknowledge Bristol-Myers Squibb for donation of the monoclonal antibody used in this work and for their financial support.

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Authors and Affiliations

  1. Department of Chemical Engineering, The Pennsylvania State University, University Park, Pennsylvania, 16802, USA

    Youngbin Baek & Andrew L. Zydney

  2. Bristol-Myers Squibb, Global Manufacturing and Supply, Devens, Massachusetts, 01434, USA

    Nripen Singh & Abhiram Arunkumar

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  1. Youngbin Baek
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  2. Nripen Singh
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  4. Andrew L. Zydney
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Correspondence to Andrew L. Zydney.

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Baek, Y., Singh, N., Arunkumar, A. et al. Effects of Histidine and Sucrose on the Biophysical Properties of a Monoclonal Antibody. Pharm Res 34, 629–639 (2017). https://doi.org/10.1007/s11095-016-2092-0

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  • DOI: https://doi.org/10.1007/s11095-016-2092-0

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