Yamada Yuma, Perez Sandra Milena Vergara, Tabata Mai, Abe Jiro, Yasuzaki Yukari, Harashima Hideyoshi
Laboratory for Molecular Design of Pharmaceutics, Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo, 060-0812, Japan.
Department of Pediatrics, Hokkaido University Hospital, Sapporo, 060-8638, Japan.
J Pharm Sci. 2015 Sep;104(9):2845-54. doi: 10.1002/jps.24310. Epub 2014 Dec 26.
The transduction of antibodies into living cells would represent a major contribution to both basic and applied biomedical fields, as currently available methods suffer from limitations such as low-uptake efficiency and endosomal entrapment. In this study, a liposome-based carrier was designed to overcome these issues. Liposomes were modified with octaarginine (R8), a cell penetrating peptide and GALA, a pH-sensitive fusogenic peptide. The presence of R8 enhanced the cellular uptake of antibodies, whereas GALA reduced endosomal entrapment, resulting in antibodies being released into the cytosol within 30 min. Moreover, compared with commercially available reagents for delivering antibodies, our system was superior in both cellular uptake and endosomal escape. In addition, specific antibodies delivered by R8-GALA liposomes were found to be associated with their epitope, confirming the preservation of functionality. This system for the efficient and high-speed cytosolic delivery of an antibody provides a valuable tool that can be useful in basic and applied research for analyzing the expression and function of intracellular molecules.
将抗体导入活细胞对基础和应用生物医学领域都将是一项重大贡献，因为目前可用的方法存在诸如摄取效率低和被内体截留等局限性。在本研究中，设计了一种基于脂质体的载体来克服这些问题。脂质体用细胞穿透肽八聚精氨酸（R8）和pH敏感的融合肽GALA进行了修饰。R8的存在增强了抗体的细胞摄取，而GALA减少了内体截留，导致抗体在30分钟内释放到细胞质中。此外，与市售的抗体递送试剂相比，我们的系统在细胞摄取和内体逃逸方面都更具优势。此外，发现由R8-GALA脂质体递送的特异性抗体与其表位相关，证实了功能的保留。这种用于高效、快速地将抗体递送至细胞质的系统提供了一种有价值的工具，可用于基础和应用研究中分析细胞内分子的表达和功能。
