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Bio‐Orthogonal, Site‐Selective Conjugation of Recombinant Proteins to Microporous Annealed Particle Hydrogels for Tissue Engineering
Author(s) -
Jivan Faraz,
Alge Daniel L.
Publication year - 2020
Publication title -
advanced therapeutics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.125
0ISSN - 2366-3987
DOI - 10.1002/adtp.201900148
Subject(s) - self healing hydrogels , recombinant dna , bone morphogenetic protein 2 , tetrazine , ethylene glycol , chemistry , mcherry , tissue engineering , green fluorescent protein , microporous material , biophysics , protein engineering , biomaterial , bone morphogenetic protein , biochemistry , biomedical engineering , combinatorial chemistry , polymer chemistry , in vitro , organic chemistry , biology , enzyme , medicine , gene
Protein conjugation to biomaterial scaffolds is a powerful approach for tissue engineering. However, typical chemical conjugation methods lack site‐selectivity and can negatively impact protein bioactivity. To overcome this problem, a site‐selective strategy is reported here for installing tetrazine groups on terminal poly‐histidines (His‐tags) of recombinant proteins. These tetrazine groups are then leveraged for bio‐orthogonal 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 His‐tag (His‐GFP), 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 protein‐2 (His‐BMP2) 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 site‐selective protein modification strategy, coupled with bio‐orthogonal 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.