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Related Experiment Videos

Nanoparticle conjugation increases protein partitioning in aqueous two-phase systems.

M Scott Long1, Christine D Keating

  • 1Department of Chemistry, Pennsylvania State University, University Park, Pennsylvania 16802, USA.

Analytical Chemistry
|January 18, 2006
PubMed
Summary

Bioconjugating proteins to gold nanoparticles significantly enhances their separation in aqueous two-phase systems (ATPS). This method offers a simple yet effective way to purify biomolecule/nanoparticle conjugates for various applications.

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Area of Science:

  • Biotechnology
  • Nanotechnology
  • Biochemistry

Background:

  • Aqueous two-phase systems (ATPS) are used for protein partitioning.
  • Controlling protein phase preference in ATPS can be challenging.
  • Biomolecule/nanoparticle conjugates have growing applications in diagnostics and materials.

Purpose of the Study:

  • To investigate the effect of bioconjugation to gold nanoparticles on protein partitioning in ATPS.
  • To explore a novel method for enhancing protein separation and purification.

Main Methods:

  • Proteins, specifically horseradish peroxidase (HRP), were conjugated to colloidal gold (Au) nanoparticles via direct adsorption.
  • The partitioning behavior of free proteins and their Au nanoparticle conjugates was analyzed in poly(ethylene glycol) (PEG)/dextran ATPS.

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  • Factors influencing partitioning, including nanoparticle size and polymer properties, were examined.

    • Main Results:

    • Free HRP showed minimal phase preference, while HRP/Au nanoparticle conjugates strongly partitioned to the PEG-rich phase (up to 150:1).
    • Other protein/Au nanoparticle conjugates demonstrated even greater partitioning (over 2000:1) to the dextran-rich phase.
    • Partitioning efficiency was influenced by nanoparticle diameter, polymer characteristics, and concentrations.

    Conclusions:

    • Bioconjugation to Au nanoparticles dramatically improves protein partitioning in ATPS without chemical modification.
    • This approach offers a facile and effective method for purifying biomolecule/nanoparticle conjugates.
    • ATPS partitioning is a powerful purification technique for conjugates used in diagnostics and materials science.