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Live-cell Imaging of Endocytic Transport using Functionalized Nanobodies in Cultured Cells
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Nanobodies for NETosis Tracking and Visualization.

Nicoleta Gutu1,2, Yuli Peng1,2, Dominik Renn1

  • 1KAUST Catalysis Center (KCC), Division of Physical Sciences & Engineering, King Abdullah University of Science and Technology, KAUST, Thuwal, Kingdom of Saudi Arabia.

Current Protocols
|October 15, 2025
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Summary
This summary is machine-generated.

Researchers developed novel nanobodies for imaging neutrophil extracellular traps (NETs) and NETosis. These engineered nanobodies targeting Ly6C/G and histones offer improved specificity and flexibility for visualizing this critical immune response.

Keywords:
NETosisVHHhistoneimagingnanobodysingle‐domain antibody

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Area of Science:

  • Immunology
  • Molecular Biology
  • Biotechnology

Background:

  • Neutrophil extracellular traps (NETs) are crucial for innate immunity, trapping pathogens.
  • NETosis, the process of NET formation, is involved in infections, inflammation, and cancer.
  • Current NETosis imaging methods have limitations in specificity and flexibility.

Purpose of the Study:

  • To develop novel nanobodies for specific and flexible imaging of NETosis.
  • To create tools for enhanced visualization of NET formation in physiological and pathological contexts.

Main Methods:

  • Engineered single-domain antibodies (nanobodies) targeting Ly6C/G and histones.
  • Optimized nanobodies for fluorescent probe labeling.
  • Utilized whole-body fluorescence tomography and intravital microscopy for visualization.

Main Results:

  • Developed Ly6C/G-targeting nanobodies for neutrophil specificity.
  • Developed anti-histone nanobodies for NET-specific targeting.
  • Demonstrated high-resolution visualization of NETosis using the engineered nanobodies.

Conclusions:

  • Modified nanobodies provide a novel and improved toolkit for NETosis imaging.
  • This approach enhances the ability to study NETosis mechanisms and therapeutic implications.
  • Offers a flexible alternative for expanding NETosis visualization capabilities.