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Related Concept Videos

Tagging and Fusion Proteins01:24

Tagging and Fusion Proteins

Proteins are involved in several cellular processes and biochemical reactions. Analyzing a specific protein of interest requires it to be isolated from the other proteins in the cell. This is achieved by overexpressing the specific gene in a suitable host to produce large quantities of the target protein. A tag or label is recombined with the gene to produce a fusion protein containing the target protein and the tag. The tags on these fusion proteins can then be used for easy detection and...

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Orthogonal Protein Purification Facilitated by a Small Bispecific Affinity Tag
10:32

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Published on: January 16, 2012

Recombinant protein purification by self-cleaving aggregation tag.

Wan-Yi Wu1, Courtney Mee, Filomena Califano

  • 1Department of Chemical Engineering, A-217 Engineering Quadrangle, Princeton University, New Jersey 08544-5263, USA.

Nature Protocols
|April 5, 2007
PubMed
Summary

This study introduces a simple, non-chromatographic method for purifying recombinant proteins using a self-cleaving tag. This elastin-like polypeptide (ELP) tag enables efficient protein isolation without complex chromatography.

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Last Updated: Jul 15, 2026

Orthogonal Protein Purification Facilitated by a Small Bispecific Affinity Tag
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Published on: April 20, 2014

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Protein Engineering

Background:

  • Recombinant protein purification is crucial for research and biotechnology.
  • Traditional chromatographic methods can be time-consuming and costly.
  • Developing simpler, scalable purification techniques is essential.

Purpose of the Study:

  • To present a novel, non-chromatographic purification technique for recombinant proteins.
  • To utilize a reversibly precipitating, self-cleaving purification tag.
  • To enable efficient isolation of pure, native target proteins.

Main Methods:

  • Employing a tripartite purification tag composed of an elastin-like polypeptide (ELP) and an intein.
  • Utilizing temperature- and salt-induced reversible precipitation of the ELP tag for purification cycles.
  • Inducing self-cleavage of the tag via a mild pH shift, followed by ELP tag precipitation.

Main Results:

  • Achieved purification of recombinant proteins expressed in Escherichia coli without chromatography.
  • Demonstrated efficient isolation of pure, native target protein after tag cleavage and precipitation.
  • Obtained high protein recoveries (50-100 mg/L) for over a dozen proteins within 8-24 hours.

Conclusions:

  • The presented method offers a simple, effective, and scalable approach for recombinant protein purification.
  • This technique eliminates the need for chromatographic separations, reducing time and cost.
  • The ELP-intein tag system provides a versatile tool for protein purification in various applications.