Electrophoretic implementation of the solution-depletion method for measuring protein adsorption, adsorption kinetics, and adsorption competition among multiple proteins in solution

Methods Mol Biol. 2013:1025:157-66. doi: 10.1007/978-1-62703-462-3_12.

Abstract

The venerable solution-depletion method is perhaps the most unambiguous method of measuring solute adsorption from solution to solid particles, requiring neither complex instrumentation nor associated interpretive theory. We describe herein an SDS-gel electrophoresis implementation of the solution--depletion method for measuring protein adsorption and protein-adsorption kinetics. Silanized-glass particles with different surface chemistry/energy and hydrophobic sepharose-based chromatographic media are used as example adsorbents. Electrophoretic separation enables quantification of adsorption competition among multiple proteins in solution for the same adsorbent surface, demonstrated herein by adsorption--competition kinetics from binary solution.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Adsorption
  • Chromatography, Agarose
  • Electrophoresis, Polyacrylamide Gel / methods*
  • Glass
  • Humans
  • Hydrophobic and Hydrophilic Interactions
  • Kinetics
  • Molecular Biology / methods
  • Proteins / chemistry*
  • Proteins / metabolism
  • Serum Albumin / chemistry
  • Solutions / chemistry*
  • Surface Properties
  • Water / chemistry

Substances

  • Proteins
  • Serum Albumin
  • Solutions
  • Water