The Protein Journal

, Volume 31, Issue 8, pp 703–709 | Cite as

Quantitation of pH-induced Aggregation in Binary Protein Mixtures by Dielectric Spectroscopy

  • Brett L. Mellor
  • Stephen J. Wood
  • Brian A. Mazzeo
Article

Abstract

This paper presents a quantitative approach for measuring pH-controlled protein aggregation using dielectric spectroscopy. The technique is demonstrated through two aggregation experiments, the first between β-lactoglobulin (β-Lg) and hen lysozyme (HENL) and the second between bovine serum albumin (BSA) and HENL. In both experiments, the formation of aggregates is strongly dependent on the solution pH and is clearly indicated by a decrease in the measured permittivity when the second protein is added. A quantifiable lower-bound on the ratio of proteins involved in the aggregation process is obtained from the permittivity spectra. Lower-bound aggregation ratios of 83 % for β-Lg/HENL at pH 6.0 and 48 % for BSA/HENL at pH 9.2 were consistent with turbidity measurements made on the same solutions.

Keywords

Dielectric spectroscopy Protein aggregation pH 

Abbreviations

β-Lg

Beta-lactoglobulin

BSA

Bovine serum albumin

EP

Electrode polarization

DI

De-ionized

DS

Dielectric spectroscopy

HENL

Hen lysozyme

PDB

Protein data bank

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Copyright information

© Springer Science+Business Media New York 2012

Authors and Affiliations

  • Brett L. Mellor
    • 1
  • Stephen J. Wood
    • 1
  • Brian A. Mazzeo
    • 1
  1. 1.Department of Electrical and Computer EngineeringBrigham Young UniversityProvoUSA

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