Viscosity Analysis of Dual Variable Domain Immunoglobulin Protein Solutions: Role of Size, Electroviscous Effect and Protein-Protein Interactions.
OBJECTIVE
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-Ig(TM)) 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-Ig(TM) protein solutions. The role of the electrostatic charge resulting in electroviscous effects for DVD-Ig(TM) protein was assessed by measuring zeta potential. Light scattering measurements were performed to detect protein-protein interactions affecting solution viscosity.


RESULTS
DVD-Ig(TM) 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-Ig(TM) 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-Ig(TM) protein at certain pH and ionic strength conditions.


CONCLUSIONS
Results show that more than one factor contributes to the increased viscosity of DVD-Ig(TM) protein and interplay of these factors modulates the overall viscosity behavior of the solution, especially at higher concentrations.
Department of Pharmaceutical Sciences, University of Connecticut, 69 North Eagleville Rd. Unit 3092, Storrs, Connecticut, 06269, USA. ashlesha0105@gmail.com.
Ashlesha S
AS
Raut
Department of Pharmaceutical Sciences, University of Connecticut, 69 North Eagleville Rd. Unit 3092, Storrs, Connecticut, 06269, USA. kalonia@uconn.edu.
Devendra S
DS
Kalonia
D000465:Algorithms; D000911:Antibodies, Monoclonal / Q000737:chemistry; D002021:Buffers; D006801:Humans; D006863:Hydrogen-Ion Concentration; D016756:Immunoglobulins, Intravenous / Q000737:chemistry*; D008970:Molecular Weight; D019999:Pharmaceutical Solutions; D011506:Proteins / Q000737:chemistry; D012996:Solutions; D055672:Static Electricity; D014783:Viscosity
D000911:Antibodies, Monoclonal; D002021:Buffers; D016756:Immunoglobulins, Intravenous; D019999:Pharmaceutical Solutions; D011506:Proteins; D012996:Solutions
bispecific antibody; charge; excluded volume; hydration; monoclonal antibody; protein formulation