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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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Related Experiment Video

Updated: Jun 1, 2026

High-throughput Purification of Affinity-tagged Recombinant Proteins
07:44

High-throughput Purification of Affinity-tagged Recombinant Proteins

Published on: August 26, 2012

Streamlined protein expression and purification using cleavable self-aggregating tags.

Lei Xing1, Wei Wu, Bihong Zhou

  • 1Department of Chemical Engineering, Tsinghua University, One Tsinghua Garden Road, Beijing 100084, China.

Microbial Cell Factories
|June 3, 2011
PubMed
Summary
This summary is machine-generated.

This study introduces a novel, matrix-free method for recombinant protein purification using self-aggregating peptides and inteins. This approach yields highly pure and active proteins, reducing costs and time for biotechnology applications.

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Related Experiment Videos

Last Updated: Jun 1, 2026

High-throughput Purification of Affinity-tagged Recombinant Proteins
07:44

High-throughput Purification of Affinity-tagged Recombinant Proteins

Published on: August 26, 2012

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

Orthogonal Protein Purification Facilitated by a Small Bispecific Affinity Tag

Published on: January 16, 2012

One-step Purification of Twin-Strep-tagged Proteins and Their Complexes on Strep-Tactin Resin Cross-linked With Bis(sulfosuccinimidyl) Suberate (BS3)
18:27

One-step Purification of Twin-Strep-tagged Proteins and Their Complexes on Strep-Tactin Resin Cross-linked With Bis(sulfosuccinimidyl) Suberate (BS3)

Published on: April 20, 2014

Area of Science:

  • Biotechnology
  • Protein Biochemistry
  • Molecular Biology

Background:

  • Recombinant protein expression and purification are critical challenges in biotechnology.
  • Self-assembling amphipathic peptides (18A and ELK16) were previously shown to induce active protein aggregates in E. coli.
  • This study builds upon prior findings to develop an improved protein purification strategy.

Purpose of the Study:

  • To develop a novel, facile, and matrix-free approach for protein expression and purification.
  • To explore the use of cleavable self-aggregating tags for streamlined protein isolation.
  • To demonstrate the efficacy of this method using various target proteins.

Main Methods:

  • Utilized cleavable self-aggregating tags composed of amphipathic peptides (18A or ELK16) and an intein.
  • Expressed target proteins as active protein aggregates in E. coli.
  • Separated aggregates via centrifugation and released proteins using intein-mediated cleavage.

Main Results:

  • Successfully purified three target proteins (lipase A, amadoriase II, β-xylosidase) with high activity and purity (>90% for intein-ELK16 fusions).
  • Achieved yields comparable to established purification methods (1.6-10.4 μg/mg wet cell pellet).
  • Demonstrated a streamlined, single-step purification process.

Conclusions:

  • The developed single-step purification method effectively produces proteins in high quantity and purity.
  • This approach significantly reduces costs and processing time.
  • Offers potential applications for both industrial scale-up and laboratory use.