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Hydrophobic Interaction Chromatography.

Brendan F O'Connor1,2,3, Philip M Cummins4

  • 1School of Biotechnology, Dublin City University, Dublin 9, Republic of Ireland. brendan.oconnor@dcu.ie.

Methods in Molecular Biology (Clifton, N.J.)
|October 13, 2016
PubMed
Summary
This summary is machine-generated.

Hydrophobic Interaction Chromatography (HIC) separates proteins using their surface hydrophobic patches. This technique is effective for protein purification in complex mixtures under mild conditions.

Keywords:
Biological activityButyl sepharoseChaotropic agentsDetergentsGlycerol gradientsHydrophobic amino acidsNonpolarOctyl sepharosePhenyl sepharose

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Area of Science:

  • Biochemistry
  • Chromatography

Background:

  • Proteins and large polypeptides possess surface hydrophobic regions composed of nonpolar amino acid side chains.
  • These hydrophobic patches are interspersed with hydrophilic regions, varying in number, size, and distribution among proteins.

Purpose of the Study:

  • To explain the principle of Hydrophobic Interaction Chromatography (HIC) for protein separation.
  • To highlight the utility of HIC in purifying proteins from complex biological mixtures.

Main Methods:

  • Hydrophobic Interaction Chromatography (HIC) exploits the differential hydrophobicity of proteins for separation.
  • The technique utilizes the presence of hydrophobic patches on protein surfaces.

Main Results:

  • HIC allows for the separation of proteins based on their surface hydrophobic characteristics.
  • The method is effective even in complex biological mixtures.

Conclusions:

  • Hydrophobic Interaction Chromatography is a widely used and effective technique for protein and large polypeptide purification.
  • The mild, protein-friendly conditions of HIC ensure good biological recoveries and maintain protein integrity.