Biomacromolecules. 2018 Nov 12;19(11):4463-4471. doi: 10.1021/acs.biomac.8b01355. Epub 2018 Oct 31.
Well-defined polymer-protein bioconjugates are widely used in therapeutics and biocatalysis. One of the challenges in the synthesis of bioconjugates is the efficient separation of the target conjugate molecules from reaction systems. In this research, surface coassembly of polymer brushes and polymer-protein bioconjugates is investigated, and it is demonstrated that the coassembly approach can be applied in the purification of polymer-protein bioconjugates. Bovine serum albumin-poly( N-isopropylacrylamide) (BSA-PNIPAM) bioconjugates were synthesized by the "grafting from" approach, and PNIPAM brushes on silica particles were prepared by the "grafting to" approach. PNIPAM brushes on silica particles are able to coassemble with BSA-PNIPAM at a temperature above the lower critical solution temperature of PNIPAM. Two-layer surface structures with collapsed PNIPAM in the inner layers and BSA in the outer layers are formed on the silica particles. The size of the silica particles and molecular weight of PNIPAM on the bioconjugates exert influences on the coassembly. The coassembly approach can be used in the purification of bioconjugates. After repeated coassembly centrifugation-release cycles, all the BSA-PNIPAM bioconjugates can be removed from the reaction solutions, and the purified bioconjugates are obtained.
结构明确的聚合物-蛋白质生物缀合物被广泛应用于治疗和生物催化。在生物缀合物的合成中,面临的挑战之一是如何从反应体系中高效分离目标缀合分子。本研究探讨了聚合物刷和聚合物-蛋白质生物缀合物的表面共组装,并证实了该共组装方法可用于聚合物-蛋白质生物缀合物的纯化。通过“从”方法合成牛血清白蛋白-聚(N-异丙基丙烯酰胺)(BSA-PNIPAM)生物缀合物,并通过“接枝到”方法制备硅胶粒子上的 PNIPAM 刷。在高于 PNIPAM 下临界溶液温度的温度下,硅胶粒子上的 PNIPAM 刷能够与 BSA-PNIPAM 共组装。在硅胶粒子上形成了内层塌陷的 PNIPAM 和外层 BSA 的双层表面结构。硅胶粒子的大小和生物缀合物上的 PNIPAM 分子量对共组装有影响。共组装方法可用于生物缀合物的纯化。经过反复的共组装离心-释放循环,可以从反应溶液中去除所有的 BSA-PNIPAM 生物缀合物,并得到纯化的生物缀合物。