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Viscosity Analysis of Dual Variable Domain Immunoglobulin Protein Solutions: Role of Size, Electroviscous Effect and Protein-Protein Interactions

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ABSTRACT

Purpose

Increased solution viscosity results in difficulties in manufacturing and delivery of therapeutic protein formulations, increasing both the time and production costs, and leading to patient inconvenience. The solution viscosity is affected by the molecular properties of both the solute and the solvent. The purpose of this work was to investigate the effect of size, charge and protein-protein interactions on the viscosity of Dual Variable Domain Immunoglobulin (DVD-IgTM) protein solutions.

Methods

The effect of size of the protein molecule on solution viscosity was investigated by measuring intrinsic viscosity and excluded volume calculations for monoclonal antibody (mAb) and DVD-IgTM protein solutions. The role of the electrostatic charge resulting in electroviscous effects for DVD-IgTM protein was assessed by measuring zeta potential. Light scattering measurements were performed to detect protein-protein interactions affecting solution viscosity.

Results

DVD-IgTM protein exhibited significantly higher viscosity compared to mAb. Intrinsic viscosity and excluded volume calculations indicated that the size of the molecule affects viscosity significantly at higher concentrations, while the effect was minimal at intermediate concentrations. Electroviscous contribution to the viscosity of DVD-IgTM protein varied depending on the presence or absence of ions in the solution. In buffered solutions, negative k D and B 2 values indicated the presence of attractive interactions which resulted in high viscosity for DVD-IgTM protein at certain pH and ionic strength conditions.

Conclusions

Results show that more than one factor contributes to the increased viscosity of DVD-IgTM protein and interplay of these factors modulates the overall viscosity behavior of the solution, especially at higher concentrations.

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Abbreviations

DLS:

Dynamic light scattering

DVD-IgTM protein:

Dual variable domain immunoglobulin protein

LLPS:

Liquid-liquid phase separation

mAb:

Monoclonal antibody

PPI:

Protein-protein Interactions

SC:

Subcutaneous

SLS:

Static light scattering

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ACKNOWLEDGMENTS AND DISCLOSURES

Authors would like to thank AbbVie Bioresearch Center, Worcester, MA for material and financial support for the work.

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences, University of Connecticut, 69 North Eagleville Rd. Unit 3092, Storrs, Connecticut, 06269, USA

    Ashlesha S. Raut & Devendra S. Kalonia

Authors
  1. Ashlesha S. Raut
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  2. Devendra S. Kalonia
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Correspondence to Ashlesha S. Raut or Devendra S. Kalonia.

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ESM 1

A brief description of T cloud as important parameters to measure PPI in solution is provided in the supporting material. Plot of T cloud for DVD-IgTM as a function of solution conditions is also elaborated. Excluded volume effect using modified Ross and Minton equation for mAb and DVD-IgTM is briefly discussed in the Supporting Information. (DOCX 146 kb)

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Raut, A.S., Kalonia, D.S. Viscosity Analysis of Dual Variable Domain Immunoglobulin Protein Solutions: Role of Size, Electroviscous Effect and Protein-Protein Interactions. Pharm Res 33, 155–166 (2016). https://doi.org/10.1007/s11095-015-1772-5

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  • DOI: https://doi.org/10.1007/s11095-015-1772-5

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