Chemically cross-linked hydrogels from repetitive protein arrays

Rossana Boni, Elizabeth A Blackburn, Dirk-Jan Kleinjan, Mantas Jonaitis, Flora Hewitt-Harris, Megan Murdoch, Susan Rosser, David C Hay, Lynne Regan

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

Biomaterials for tissue regeneration must mimic the biophysical properties of the native physiological environment. A protein engineering approach allows the generation of protein hydrogels with specific and customised biophysical properties designed to suit a particular physiological environment. Herein, repetitive engineered proteins were successfully designed to form covalent molecular networks with defined physical characteristics able to sustain cell phenotype. Our hydrogel design was made possible by the incorporation of the SpyTag (ST) peptide and multiple repetitive units of the SpyCatcher (SC) protein that spontaneously formed covalent crosslinks upon mixing. Changing the ratios of the protein building blocks (ST:SC), allowed the viscoelastic properties and gelation speeds of the hydrogels to be altered and controlled. The physical properties of the hydrogels could readily be altered further to suit different environments by tuning the key features in the repetitive protein sequence. The resulting hydrogels were designed with a view to allow cell attachment and encapsulation of liver derived cells. Biocompatibility of the hydrogels was assayed using a HepG2 cell line constitutively expressing GFP. The cells remained viable and continued to express GFP whilst attached or encapsulated within the hydrogel. Our results demonstrate how this genetically encoded approach using repetitive proteins could be applied to bridge engineering biology with nanotechnology creating a level of biomaterial customisation previously inaccessible.

Original languageEnglish
Article number107981
Number of pages8
JournalJournal of Structural Biology
Issue number3
Early online date26 May 2023
Publication statusPublished - 1 Sept 2023

Keywords / Materials (for Non-textual outputs)

  • protein engineering
  • repetitive sequence
  • hydrogels
  • SpyTag SpyCatcher
  • protein rheology
  • tissue engineering
  • biomaterials


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