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Updated: Jan 27, 2026

Efficient Sampling of Genetically Encoded Biosensor Design Space Enabled with a Design of Experiments and Automation Workflow
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Scalable Geometrically Designed Protein Cages Assembled via Genetically Encoded Split Inteins.

James N Wright1, Wan Ling Wong1, Joseph A Harvey1

  • 1School of Biological and Chemical Sciences, Queen Mary, University of London, Mile End Road, London E1 4NS, UK.

Structure (London, England : 1993)
|March 19, 2019
PubMed
Summary

Researchers developed a versatile protein assembly system using split intein ligation to create scalable protein cages. This method allows for efficient, one-pot or stepwise construction of complex protein structures for biotechnological applications.

Keywords:
directed protein assemblynative chemical ligationprotein conjugationprotein designprotein semi-synthesissplit intein ligation

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Area of Science:

  • Biochemistry
  • Structural Biology
  • Protein Engineering

Background:

  • Protein self-assembly is crucial for biotechnological applications.
  • Designing generic protein assembly systems remains a significant challenge.

Purpose of the Study:

  • To develop an expandable recombinant assembly system for scalable protein cage production.
  • To enable both one-pot and stepwise construction of complex protein structures.

Main Methods:

  • Utilized split intein-mediated native chemical ligation for protein assembly.
  • Employed complementary oligomeric "half-cage" and extendable monomeric "linker" protein fusions.
  • Incorporated modular protein domains and orthogonal split intein pairs to drive assembly.

Main Results:

  • Demonstrated one-pot and stepwise assembly of protein cages using consensus-designed repeat proteins.
  • Achieved rapid, high-yielding ligations under mild conditions.
  • Successfully purified discrete protein cages forming trigonal bipyramidal structures.

Conclusions:

  • The developed system offers a versatile and scalable approach for engineering protein cages.
  • This method facilitates the creation of user-defined protein structures for diverse biotechnological uses.