Multiphoton lithography with protein photoresists

Dmitry Sivun, Eljesa Murtezi, Tina Karimian, Kurt Hurab, Maryam Marefat, Elena Klimareva, Christoph Naderer, Boris Buchroithner, Thomas a. Klar, Georgii Gvindzhiliia, Andreas Horner, Jaroslaw Jacak

Research output: Contribution to journalArticlepeer-review


Recently, 2D/3D direct laser writing has attracted increased attention due to its broad applications ranging from biomedical engineering to aerospace. 3D nanolithography of water-soluble protein-based scaffolds have been envisioned to provide a variety of tunable properties. In this paper, we present a functional protein-based photoresist with tunable mechanical properties that is suitable for multiphoton lithography (MPL). Through the use of methacrylated streptavidin or methacrylated bovine serum albumin in combination with polyethylene glycol diacrylate or methacrylated hyaluronic acid as crosslinkers and a vitamin-based photoinitiator, we were able to write two- and three-dimensional structures as small as 200 nm/600 nm lateral/axial features, respectively. We also demonstrated that Young's modulus can be tuned by the photoresist composition, and we were able to achieve values as low as 40 kPa. Furthermore, we showed that Young's modulus can be recovered after drying and rehydration (i.e. shelf time determination). The retained biological functionality of the streptavidin scaffolds was demonstrated using fluorescently labelled biotins. Using single-molecule fluorescence microscopy, we estimated the density of streptavidin in the written features (1.8 ± 0.2 × 10 5 streptavidins per 1.00 ± 0.05 μm³ of feature volume). Finally, we showed applicability of our 2D scaffold as a support for a fluorescence absorbance immuno-assay (FLISA), and as a delivery platform of extracellular vesicles to HeLa cells.

Original languageEnglish
Article number100994
Pages (from-to)100994
JournalMaterials Today Bio
Publication statusPublished - Apr 2024


  • Extracellular vesicle
  • Functional photoresist
  • Multiphoton lithography
  • Protein printing
  • Tissue scaffolds


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