South China Advanced Institute for Soft Matter Science and Technology, School of Molecular Science and Engineering, South China University of Technology, Guangzhou 510640, China.
Biomater Sci. 2020 Jul 7;8(13):3741-3750. doi: 10.1039/d0bm00679c. Epub 2020 Jun 5.
Cytosolic protein delivery technique plays an important role in protein-based biotechnologies and therapeutics. However, the development of efficient nanocarriers for delivering cargo proteins into cytosols remains a continuing challenge due to the existence of multiple barriers. Here, we report an efficient strategy for the cytosolic delivery of native proteins by surface-engineered gold nanoparticles combined with hypertonicity treatment. Sub-10 nm gold nanoparticles stabilized by both cysteamine and 4-mercaptophenylboronic acid were used to complex cargo proteins via a combination of nitrogen-boronate coordination and ionic interactions. The yielding protein complexes with a size around 100 nm showed efficient endocytosis via micropinocytosis- and lipid raft-mediated pathways. Further the hypertonicity treatment of the transduced cells by glycerol, glucose, sucrose, and NaCl solutions efficiently facilitates the endosomal escape and the intracellular release of cargo proteins. By the proposed strategy, cargo proteins including bovine serum albumin, ovalbumin, green fluorescent protein, R-phycoerythrin, and horseradish peroxidase were successfully delivered into cell cytosol with maintained protein bioactivity. This study provides a feasible and efficient strategy for the intracellular protein delivery.
细胞质蛋白递送技术在蛋白质生物技术和治疗中起着重要作用。然而，由于存在多种障碍，开发将货物蛋白递送至细胞质的有效纳米载体仍然是一个持续的挑战。在这里，我们报告了一种通过表面工程金纳米粒子结合高渗处理实现天然蛋白质细胞质内有效递送的策略。通过氮硼酸配位和离子相互作用的组合，使用同时由半胱氨酸和 4-巯基苯硼酸稳定的亚 10nm 金纳米粒子来复合货物蛋白。尺寸约为 100nm 的产率蛋白复合物通过微绒毛和脂筏介导的途径有效内化。此外，甘油、葡萄糖、蔗糖和 NaCl 溶液对转导细胞的高渗处理可有效促进内体逃逸和货物蛋白的细胞内释放。通过所提出的策略，包括牛血清白蛋白、卵清蛋白、绿色荧光蛋白、藻红蛋白和辣根过氧化物酶在内的货物蛋白被成功递送至细胞细胞质内，且保持了蛋白生物活性。这项研究为细胞内蛋白递送提供了一种可行且高效的策略。
