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Cell-Compatible, Site-Specific Covalent Modification of Hydrogel Scaffolds Enables User-Defined Control over Cell-Material Interactions

Biomacromolecules. 2019-07; 
HammerJoshua A,RutaAnna,TherienAidan M,WestJennif
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Gene Synthesis … Partial gene constructs containing the N-terminal sequence (RGDS or RDGS), mCherry, and the 6xHis tag complex were purchased from Genscript as inserts within the pGS … the gene for SpyCatcher was purchased from IDT DNA and then inserted into the Genscript … Monoacrylate-PEG-SpyTag was synthesized as follows (Figure 1a). SpyTag peptide (AHIVMVDAYKPTK) was purchased from GenScript with an added N-terminal cysteine bearing a thiol group, dubbed C-SpyTag. Get A Quote
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摘要

SpyCatcher, a 15 kDa protein domain that spontaneously forms a site-specific covalent bond with the 13 amino acid peptide SpyTag, was used to covalently link a model recombinant protein containing a SpyCatcher domain and the adhesive ligand Arg-Gly-Asp-Ser (RGDS) (RGDS-SC) into SpyTag-containing poly(ethylene glycol) (PEG) hydrogels. This new strategy for covalent immobilization of proteins or peptides provides an easy and gentle mechanism for biochemical modification of hydrogels. Labeling efficiency was approximately 100% when soluble RGDS-SC was applied to SpyTag-containing hydrogels at a 1:1 molar ratio. RGDS-SC remained stably bound throughout the 5 days of rinsing, and 3T3 fibroblasts were able to a... More

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