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Sortase A-mediated multi-functionalization of protein nanoparticles.

Qi Chen1, Qing Sun, Nicholas M Molino

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We developed a fast method for modifying protein nanoparticles using Sortase A ligation. This technique allows for easy purification and maintains nanoparticle integrity for applications in biocatalysis, biosensing, and imaging.

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Area of Science:

  • Bioconjugation Chemistry
  • Nanobiotechnology
  • Protein Engineering

Background:

  • Protein nanoparticles offer versatile platforms for various applications.
  • Efficient and site-specific functionalization methods are crucial for controlling nanoparticle properties.
  • Sortase A-mediated ligation is a powerful tool for protein modification.

Purpose of the Study:

  • To develop a rapid, covalent, and site-directed functionalization strategy for protein nanoparticles.
  • To demonstrate the applicability of this method across a range of protein sizes.
  • To facilitate easy purification and ensure the stability of functionalized nanoparticles.

Main Methods:

  • Utilizing Sortase A-mediated ligation for covalent attachment of functional proteins to nanoparticles.
  • Employing a thermo-responsive elastin-like-peptide for facile nanoparticle purification.
  • Characterizing the integrity and activity of modified nanoparticles after purification and phase transitions.

Main Results:

  • Successfully achieved fast, covalent, and site-directed functionalization of protein nanoparticles.
  • Demonstrated successful modification of monomeric to large tetrameric protein structures.
  • Obtained easy purification of modified E2 nanoparticles via elastin-like-peptide functionalization.
  • Confirmed that protein nanoparticles remained intact and active after repeated thermo-responsive phase transitions.

Conclusions:

  • The developed Sortase A ligation strategy provides an efficient method for protein nanoparticle functionalization.
  • The thermo-responsive peptide tag enables straightforward purification, enhancing the practical utility of modified nanoparticles.
  • These robust and active protein nanoparticles are suitable for advanced applications in biocatalysis, biosensing, and imaging.