Effect of Aggregation on the Hydrodynamic Properties of Bovine Serum Albumin

  • Mariya A. Pindrus
  • James L. Cole
  • Japneet Kaur
  • Steven J. Shire
  • Sandeep Yadav
  • Devendra S. Kalonia
Research Paper
  • 86 Downloads

Abstract

Purpose

To systematically analyze shape and size of soluble irreversible aggregates and the effect of aggregate formation on viscosity.

Methods

Online light scattering, refractive index and viscosity detectors attached to HPLC (Viscotek®) were used to study aggregation, molecular weight and intrinsic viscosity of bovine serum albumin (BSA). Irreversible aggregates were generated by heat stress. Bulk viscosity was measured by an oscillating piston viscometer.

Results

As BSA was heated at a higher concentration or for a longer time, the relative contribution, molecular weight and intrinsic viscosity of aggregate species increased. Molecular shape was evaluated from intrinsic viscosity values, and aggregates were estimated to be more asymmetric than monomer species. The presence of aggregates resulted in an increase in bulk viscosity when relative contribution of very high molecular weight species exceeded 10%.

Conclusions

For model system and conditions studied, generation of higher order aggregate species was concluded to be associated with an increase in molecular asymmetry. Elevated viscosity in the presence of aggregated species points to molecular asymmetry being a critical parameter affecting solution viscosity of BSA.

KEY WORDS

aggregation effective molecular shape intrinsic viscosity 

ABBREVIATIONS

BSA

Bovine serum albumin

DP

Differential pressure

HMW

High molecular weight

LALS

Low angle light scattering

mAb

Monoclonal antibody

mM

Millimolar

PPI

Protein-protein interactions

RALS

Right angle light scattering

RI

Refractive index

SEC

Size exclusion chromatography

TDA

Triple detector array

vHMW

Very high molecular weight

Supplementary material

11095_2017_2231_MOESM1_ESM.docx (81 kb)
ESM 1(DOCX 81 kb)

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Copyright information

© Springer Science+Business Media, LLC 2017

Authors and Affiliations

  • Mariya A. Pindrus
    • 1
  • James L. Cole
    • 2
  • Japneet Kaur
    • 1
  • Steven J. Shire
    • 3
  • Sandeep Yadav
    • 3
  • Devendra S. Kalonia
    • 1
  1. 1.Department of Pharmaceutical SciencesUniversity of ConnecticutStorrsUSA
  2. 2.Department of Molecular and Cell Biology and Department of ChemistryUniversity of ConnecticutStorrsUSA
  3. 3.Late Stage Pharmaceutical DevelopmentGenentech, Inc.South San FranciscoUSA

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