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A Method of Targeted Cell Isolation via Glass Surface Functionalization
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Published on: September 20, 2016

Sortase A as a tool to functionalize surfaces.

Tjitske Sijbrandij1, Nivedita Cukkemane, Kamran Nazmi

  • 1Department of Oral Biochemistry, Academic Centre for Dentistry Amsterdam, University of Amsterdam and VU University Amsterdam, Gustav Mahlerlaan 3004, 1081 LA Amsterdam, The Netherlands.

Bioconjugate Chemistry
|March 28, 2013
PubMed
Summary
This summary is machine-generated.

This study demonstrates enzymatic surface functionalization using Sortase A (SrtA) to attach poly(ethylene glycol) (PEG). This method effectively reduced bacterial adherence, offering a novel approach to combat surface fouling.

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Area of Science:

  • Biomaterials Engineering
  • Surface Chemistry
  • Biotechnology

Background:

  • Surface fouling by biofoulants is a significant challenge in various applications.
  • Functionalization with poly(ethylene glycol) (PEG) is a common strategy to prevent biofouling.
  • Enzymatic methods offer precise control for surface modification.

Purpose of the Study:

  • To establish a proof of concept for enzymatically coupling PEG to peptide-functionalized surfaces.
  • To utilize the enzyme Sortase A (SrtA) for site-specific PEG conjugation.
  • To evaluate the efficacy of this method in reducing bacterial adhesion.

Main Methods:

  • Hydrophobic polystyrene surfaces were coated with an anchoring peptide (P3) containing a pentaglycine motif for SrtA recognition.
  • Transpeptidation was performed using SrtA to conjugate either biotin or PEG, both equipped with the sortase recognition motif (LPETG).
  • Biotinylation levels were quantified to confirm SrtA activity, and bacterial adherence assays were conducted on PEG-ylated surfaces.

Main Results:

  • High levels of surface-bound biotin were observed when using biotin-LPETG and SrtA, confirming successful enzymatic conjugation.
  • Minimal reactivity was detected with a scrambled SrtA motif or in the absence of SrtA.
  • Conjugation of PEG to the surface significantly decreased bacterial adherence.

Conclusions:

  • Sortase A (SrtA) can be effectively employed for the enzymatic coupling of poly(ethylene glycol) (PEG) to peptide-modified surfaces.
  • This enzymatic approach provides a robust method for creating antifouling surfaces.
  • The developed PEGylation strategy significantly reduces bacterial adhesion, demonstrating its potential for combating surface fouling.