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A Femtoliter Droplet Array for Massively Parallel Protein Synthesis from Single DNA Molecules
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Multiphoton lithography with protein photoresists.

Dmitry Sivun1, Eljesa Murtezi1, Tina Karimian1

  • 1Department of Medical Engineering, University of Applied Sciences Upper Austria, Garnisonstraße 21, 4020, Linz, Austria.

Materials Today. Bio
|February 22, 2024
PubMed
Summary

Researchers developed a novel protein-based photoresist for 2D/3D direct laser writing. This material allows tunable mechanical properties and retains biological function for applications in tissue engineering and diagnostics.

Keywords:
Extracellular vesicleFunctional photoresistMultiphoton lithographyProtein printingTissue scaffolds

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Area of Science:

  • Materials Science
  • Biotechnology
  • Nanotechnology

Background:

  • Direct laser writing (DLW) is a powerful nanofabrication technique.
  • Protein-based materials offer biocompatibility and tunable properties for advanced applications.
  • Existing protein-based photoresists often lack control over mechanical properties and long-term stability.

Purpose of the Study:

  • To develop a novel protein-based photoresist for multiphoton lithography (MPL).
  • To achieve tunable mechanical properties in 2D and 3D protein scaffolds.
  • To demonstrate the biological functionality and potential applications of the developed material.

Main Methods:

  • Formulation of a protein-based photoresist using methacrylated proteins (streptavidin or bovine serum albumin) and crosslinkers (polyethylene glycol diacrylate or methacrylated hyaluronic acid) with a vitamin-based photoinitiator.
  • Fabrication of 2D and 3D structures using multiphoton lithography (MPL).
  • Characterization of mechanical properties (Young's modulus) and biological functionality (streptavidin-biotin binding) using techniques like single-molecule fluorescence microscopy.

Main Results:

  • Successfully fabricated 2D and 3D protein structures with feature sizes down to 200 nm (lateral) and 600 nm (axial).
  • Demonstrated tunable Young's modulus from 40 kPa, with recovery after drying and rehydration, indicating shelf-life stability.
  • Confirmed retained biological functionality of streptavidin scaffolds and quantified streptavidin density (1.8 × 10^5 per μm³).
  • Showcased applicability in a fluorescence absorbance immunoassay (FLISA) and as a delivery platform for extracellular vesicles.

Conclusions:

  • A versatile protein-based photoresist for MPL with tunable mechanical properties and preserved biological activity has been developed.
  • The material enables the fabrication of high-resolution 2D/3D scaffolds suitable for biomedical applications, including diagnostics and drug/vesicle delivery.
  • This work opens new avenues for advanced protein-based nanomaterials in tissue engineering and personalized medicine.