A novel alternative was developed for the synthesis of polymer-protein conjugates with a controllable number of polymer chains. Human serum albumin (HSA), as a model protein, was grafted by poly(N-isopropyl-acrylamide) (PNIPAM) using N-succinimidyl-3-(2-pyridyldithiol) propionate as a coupling agent at room temperature in aqueous media. The influence of grafting number on the stability of the protein-polymer conjugate against the degradation of enzyme was examined, and the temperature dependent bioactivity was monitored through testing the warfarin-binding affinity. Furthermore, the protein could be recovered by incubating the conjugate under reducing conditions to cause the cleavage of the multiple disulfide bonds. © The Royal Society of Chemistry 2014.
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