Covalent Modification of Synthetic Hydrogels with Bioactive Proteins via Sortase-Mediated Ligation
Author(s)Cambria, Elena; Renggli, Kasper; Kroll, Carsten; Krueger, Andrew T.; Imperiali, Barbara; Griffith, Linda G.; Chopko Ahrens, Caroline; Cook, Christi Dionne; ... Show more Show less
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Synthetic extracellular matrices are widely used in regenerative medicine and as tools in building in vitro physiological culture models. Synthetic hydrogels display advantageous physical properties, but are challenging to modify with large peptides or proteins. Here, a facile, mild enzymatic postgrafting approach is presented. Sortase-mediated ligation was used to conjugate human epidermal growth factor fused to a GGG ligation motif (GGG-EGF) to poly(ethylene glycol) (PEG) hydrogels containing the sortase LPRTG substrate. The reversibility of the sortase reaction was then exploited to cleave tethered EGF from the hydrogels for analysis. Analyses of the reaction supernatant and the postligation hydrogels showed that the amount of tethered EGF increases with increasing LPRTG in the hydrogel or GGG-EGF in the supernatant. Sortase-tethered EGF was biologically active, as demonstrated by stimulation of DNA synthesis in primary human hepatocytes and endometrial epithelial cells. The simplicity, specificity, and reversibility of sortase-mediated ligation and cleavage reactions make it an attractive approach for modification of hydrogels.
DepartmentMassachusetts Institute of Technology. Center for Gynepathology Research; Massachusetts Institute of Technology. Department of Biological Engineering; Massachusetts Institute of Technology. Department of Biology; Massachusetts Institute of Technology. Department of Chemical Engineering; Massachusetts Institute of Technology. Department of Chemistry
American Chemical Society (ACS)
Cambria, Elena, Kasper Renggli, Caroline C. Ahrens, Christi D. Cook, Carsten Kroll, Andrew T. Krueger, Barbara Imperiali, and Linda G. Griffith. “Covalent Modification of Synthetic Hydrogels with Bioactive Proteins via Sortase-Mediated Ligation.” Biomacromolecules 16, no. 8 (August 10, 2015): 2316–26. © 2015 American Chemical Society
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