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Aggregating tags for column-free protein purification.

Zhanglin Lin1, Qing Zhao2, Lei Xing2

  • 1Department of Chemical Engineering, National Engineering Laboratory for Industrial Enzymes, Tsinghua University, Beijing, China. zhanglinlin@mail.tsinghua.edu.cn.

Biotechnology Journal
|November 12, 2015
PubMed
Summary

New aggregating tags offer a cost-effective, column-free method for recombinant protein purification, achieving purity comparable to His-tag. These tags induce protein aggregation, simplifying separation from impurities.

Keywords:
Aggregating tagsCleavable tags /Column-freeProtein purificationPurification tags

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Area of Science:

  • Biotechnology
  • Protein Chemistry
  • Molecular Biology

Background:

  • Protein purification is essential for biotechnology.
  • Conventional methods like His-tag purification can be costly and time-consuming.
  • Aggregating tags offer a promising alternative for column-free protein purification.

Purpose of the Study:

  • To review and categorize emerging aggregating tags for protein purification.
  • To compare the advantages and disadvantages of aggregating tags with conventional purification methods.
  • To assess the potential of aggregating tags for cost-effective recombinant protein production.

Main Methods:

  • Categorization of aggregating tags based on their aggregation behavior in vivo and in vitro.
  • Discussion of the advantages and disadvantages of each tag category.
  • Comparison of aggregating tags with traditional tags such as His-tag, maltose-binding protein (MBP) tag, and intein-mediated purification with a chitin-binding tag (IMPACT-CN).

Main Results:

  • Aggregating tags provide a column-free purification alternative with yields and purity comparable to His-tag.
  • Tags are categorized into those inducing inactive aggregates in vivo, active aggregates in vivo, or soluble expression with in vitro aggregation.
  • While promising, further systematic testing is needed to optimize the use of these tags.

Conclusions:

  • Aggregating tags represent a significant advancement in cost-effective protein purification.
  • These tags can simplify recombinant protein production, especially at industrial scales.
  • Combining aggregating tags with traditional methods may further reduce purification costs by minimizing resin and column expenses.