Protein polymer hybrids represent an important and rapidly expanding class of biomaterials. Typically in these hybrids the linkage between the protein and the polymer is covalent. Methods for synthesis of these biohybrids can be generally sorted into the grafting-from or grafting-to strategy. These strategies are thought to be interchangeable with the only major decisions being a choice of synthetic accessibility and purification of the desired hybrid. In our study we found that the choice of the polymer ligation strategy played a major role in the stability and bioactivity of the final hybrid. We have chosen DNase I as a model RE for modification due to its potential clinical and pharmacological relevance. DNase I degrades DNA to lower molecular weight fragments through the catalysis of non-specific hydrolysis of double-stranded DNA predominantly. It is applied in preparation of RNA for a variety of purposes, including gene expression analysis and exome sequencing. In addition, DNase I is used to induce bacterial susceptibility to antibiotics and removal of biofilms from surfaces through the degradation of extracellular-DNA. Herein we found that the grafting-from strategy using either reversible addition-fragmentation chain transfer polymerization (RAFT) or atom transfer radical polymerization (ATRP) yielded biohybrids with no activity due to nonspecific reaction of initiator radical with amino acids on protein surface, whereas the grafting-to method gave active, thermally stable biohybrids.

Importance of polymer grafting strategy for the retention of DNase I polymer hybrid activity

256th National Meeting (2018)

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