The specific delivery of proteins to human liver cells by engineered bio-nanocapsules
The specific delivery of proteins to human liver cells by engineered bio-nanocapsules
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DOI：
10.1111/j.1742-4658.2005.04790.x
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发表时间：
2005-07-01
期刊：
影响因子：
5.4
通讯作者：
Seno, M
中科院分区：
文献类型：
--
作者：
Yu, DW;Amano, C;Seno, M
由B型肝炎病毒表面抗原（sAg）组成的生物纳米胶囊（BNC）是一种高效的纳米机器，可实现基因和药物的肝脏特异性递送。大约110个sAg分子结合形成平均直径为130 nm的BNC颗粒。L蛋白是主要由preS和S区组成的sAg肽。前S区，与人类肝细胞的特异性亲和力，是本地化的N-末端。preS之后的S区具有两个负责形成颗粒的跨膜区。在这项研究中，祖母绿绿色荧光蛋白（EGFP）的融合在C-末端的S区的目的是提供蛋白质到人肝细胞。S区的C-末端的截断是必需的，以获得足够的表达水平在Cos 7细胞。所产生的纳米颗粒将EGFP递送到人类肝癌细胞，将EGFP部分展示在外部或将其封闭在内部。然而，仅单一取向表征颗粒，使得任一类型的L融合颗粒可以通过抗体亲和柱有效且独立地分离。在本研究中设计的L融合颗粒的双C-末端拓扑结构可以应用于L融合蛋白的C-末端部分的各种蛋白质，这取决于蛋白质的特性，例如细胞质蛋白，以及细胞因子或细胞表面受体的配体。我们认为这种融合设计是制备将蛋白质递送到特定细胞或组织的BNC的最有效方法。
A bio-nanocapsule (BNC), composed of the surface antigen (sAg) of the hepatitis B virus, is an efficient nanomachine with which to accomplish the liver-specific delivery of genes and drugs. Approximately 110 molecules of sAg are associated to form a BNC particle with an average diameter of 130 nm. The L protein is an sAg peptide composed mainly of preS and S regions. The preS region, with specific affinity for human hepatocytes, is localized in the N-terminus. The S region following the preS has two transmembrane regions responsible for the formation of particles. In this study, the fusion of emerald green fluorescent protein (EGFP) at the C-terminus of the S region was designed to deliver proteins to human hepatocytes. Truncation of the C-terminus of the S region was required to obtain sufficient expression levels in Cos7 cells. The nanoparticles that were produced delivered EGFP to human hepatoma cells, displaying the EGFP moiety outside, or enclosing it inside. However, only a single orientation characterizes the particle, so that either type of L fusion particle could be effectively and independently separated by an antibody affinity column. The dual C-terminal topologies of the L fusion particles designed in this study could be applied to various proteins for the C-terminal moiety of the L fusion proteins, depending on the character of the proteins, such as cytoplasmic proteins, as well as cytokines or ligands to cell surface receptors. We suggest that this fusion design is the most efficient way to prepare a BNC that delivers proteins to specific cells or tissues.