Bio-Orthogonal, Site-Selective Conjugation of Recombinant Proteins to Microporous Annealed Particle Hydrogels for Tissue Engineering Academic Article uri icon

abstract

  • AbstractProtein conjugation to biomaterial scaffolds is a powerful approach for tissue engineering. However, typical chemical conjugation methods lack siteselectivity and can negatively impact protein bioactivity. To overcome this problem, a siteselective strategy is reported here for installing tetrazine groups on terminal polyhistidines (Histags) of recombinant proteins. These tetrazine groups are then leveraged for bioorthogonal conjugation to poly(ethylene glycol) hydrogel microparticles, which are subsequently assembled into microporous annealed particle (MAP) hydrogels. The efficacy of the strategy is demonstrated using recombinant, green fluorescent protein with a Histag (HisGFP), which enhances fluorescence of the MAP hydrogels compared to control protein lacking tetrazine groups. Subsequently, to demonstrate efficacy with a therapeutic protein, recombinant human bone morphogenetic protein2 (HisBMP2) is conjugated. Human mesenchymal stem cells growing in the MAP hydrogels respond to the conjugated BMP2 and significantly increase mineralization after 21 days compared to controls. Thus, this siteselective protein modification strategy, coupled with bioorthogonal click chemistry, is expected to be useful for bone defect repair and regeneration therapies. Broader application to the integration of protein therapeutics with biomaterials is also envisioned.

published proceedings

  • ADVANCED THERAPEUTICS

altmetric score

  • 1.5

author list (cited authors)

  • Jivan, F., & Alge, D. L.

citation count

  • 5

publication date

  • January 2020

publisher