Electrophoretic Implementation of the Solution-Depletion Method for Measuring Protein Adsorption, Adsorption Kinetics, and Adsorption Competition Among Multiple Proteins in Solution

  • Hyeran Noh
  • Naris Barnthip
  • Purnendu Parhi
  • Erwin A. Vogler
Part of the Methods in Molecular Biology book series (MIMB, volume 1025)


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.

Key words

Protein adsorption Adsorption kinetics Adsorption competition Electrophoresis Solution depletion 



This work was supported by National Institute of Health grant PHS 5R01HL069965. The authors appreciate the support from the Departments of Materials Science and Engineering and Bioengineering, The Pennsylvania State University.


  1. 1.
    Vogler EA (2012) Protein adsorption in three dimensions. Biomaterials 33:1201–1237CrossRefGoogle Scholar
  2. 2.
    Anderson NL et al (2004) The human plasma proteome—a nonredundant list developed by combination of four separate sources. Mol Cell Proteomics 3(4):311–326CrossRefGoogle Scholar
  3. 3.
    Noh H, Vogler EA (2006) Volumetric interpretation of protein adsorption: partition coefficients, interphase volumes, and free energies of adsorption to hydrophobic surfaces. Biomaterials 27:5780–5793CrossRefGoogle Scholar
  4. 4.
    Noh H, Vogler EA (2006) Volumetric interpretation of protein adsorption: mass and energy balance for albumin adsorption to particulate adsorbents with incrementally-increasing hydrophilicity. Biomaterials 27:5801–5812CrossRefGoogle Scholar
  5. 5.
    Noh H, Vogler EA (2007) Volumetric interpretation of protein adsorption: competition from mixtures and the vroman effect. Biomaterials 28:405–422CrossRefGoogle Scholar
  6. 6.
    Noh H, Vogler EA (2008) Volumetric interpretation of protein adsorption: ion-exchange adsorbent capacity, protein pI, and interaction energetics. Biomaterials 29:2033–2048CrossRefGoogle Scholar
  7. 7.
    Barnthip N et al (2008) Volumetric interpretation of protein adsorption: kinetic consequences of a slowly-concentrating interphase. Biomaterials 29:3062–3074CrossRefGoogle Scholar
  8. 8.
    Barnthip N et al (2009) Volumetric interpretation of protein adsorption: kinetics of protein-adsorption competition from binary solution. Biomaterials 30:6495–6513CrossRefGoogle Scholar
  9. 9.
    Barnthip N, Vogler EA (2012) Protein adsorption kinetics from single- and binary-solution. Appl Surf SciGoogle Scholar
  10. 10.
    Parhi P et al (2010) Role of water and proteins in the attachment of mammalian cells to surfaces: a review. J Adhesion Sci and Tech 24:853–888CrossRefGoogle Scholar
  11. 11.
    Parhi P et al (2009) Volumetric interpretation of protein adsorption: capacity scaling with adsorbate molecular weight and aomaterials 30:6814–6824CrossRefGoogle Scholar
  12. 12.
    Kao P et al (2010) Volumetric interpretation of protein adsorption: interfacial packing of protein adsorbed to hydrophobic surfaces from surface-saturating solution concentrations. Biomaterials 32:969–978CrossRefGoogle Scholar
  13. 13.
    Sagiv J et al (1986) Self-assembling monolayers: a study of their formation, composition, and structure. In: Mittal KL, Bothorel P (eds) Surfactants in solution, vol 5. Plenum Press, New York, pp 965–978CrossRefGoogle Scholar
  14. 14.
    Fadeev AY, McCarthy TJ (2000) Self-assembly is not the only reaction possible between alkyltrichlorosilanes and surfaces: monomolecular and oligomeric covalently attached layers of dichloro- and trichloroalkylsilanes on silicon. Langmuir 16:7268–7274CrossRefGoogle Scholar
  15. 15.
    Wang M et al (2005) Self-assembled silane monolayers: fabrication with nanoscale uniformity. Langmuir 21(5):1848–1857CrossRefGoogle Scholar
  16. 16.
    Oner D, McCarthy TJ (2000) Ultrahydrophobic surfaces. Effects of topography length scales on wettability. Langmuir 165:7777–7782CrossRefGoogle Scholar
  17. 17.
    Gao L, McCarthy TJ (2009) Wetting 101. Langmuir 25(24):14105–14115CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Hyeran Noh
    • 1
  • Naris Barnthip
    • 2
  • Purnendu Parhi
    • 3
  • Erwin A. Vogler
    • 4
    • 5
  1. 1.Department of Optometry and Vision ScienceSeoul National University and TechnologySeoulSouth Korea
  2. 2.Division of PhysicsRajamangala University of TechnologyThanyaburiThailand
  3. 3.Department of ChemistryRavenshaw UniversityCuttackIndia
  4. 4.Department of Materials Science and EngineeringThe Pennsylvania State UniversityUniversity ParkUSA
  5. 5.Department of BioengineeringThe Pennsylvania State UniversityUniversity ParkUSA

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