Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Selangor, Malaysia.
J Appl Microbiol. 2012 Jan;112(1):119-31. doi: 10.1111/j.1365-2672.2011.05176.x. Epub 2011 Nov 14.
To display a liver-specific ligand on the hepatitis B virus core particles for cell-targeting delivery.
A liver cell-binding ligand (preS1) was fused at the N-terminal end of the hepatitis B core antigen (HBcAg), but the fusion protein (preS1His(6) HBcAg) was insoluble in Escherichia coli and did not form virus-like particles (VLPs). A method to display the preS1 on the HBcAg particle was established by incorporating an appropriate molar ratio of the truncated HBcAg (tHBcAg) to the preS1His(6) HBcAg. Gold immunomicroscopy showed that the subunit mixture reassembled into icosahedral particles, displaying the preS1 ligand on the surface of VLPs. Fluorescence microscopy revealed that the preS1 ligand delivered the fluorescein-labelled VLPs into the HepG2 cells efficiently.
Chimeric VLPs containing the insoluble preS1His(6) HBcAg and highly soluble tHBcAg were produced by a novel incorporation method. The preS1 ligand was exposed on the surface of the VLPs and was shown to deliver fluorescein molecules into liver cells.
The newly established incorporation method can be used in the development of chimeric VLPs that could serve as potential nanovehicles to target various cells specifically by substituting the preS1 ligand with different cell-specific ligands.
在乙型肝炎病毒核心颗粒上展示一种肝脏特异性配体，用于细胞靶向递药。
将一个肝细胞结合配体（前 S1 区）融合到乙型肝炎核心抗原（HBcAg）的 N 端，但融合蛋白（preS1His(6) HBcAg）在大肠杆菌中不溶，也不能形成病毒样颗粒（VLPs）。通过加入适当比例的截短 HBcAg（tHBcAg）到 preS1His(6) HBcAg 中，建立了一种在 HBcAg 颗粒上展示前 S1 区的方法。金免疫电镜显示，亚单位混合物重新组装成二十面体颗粒，在 VLPs 表面展示前 S1 配体。荧光显微镜显示，前 S1 配体有效地将荧光素标记的 VLPs 递送到 HepG2 细胞中。
通过一种新的掺入方法生产了含有不溶性 preS1His(6) HBcAg 和高可溶性 tHBcAg 的嵌合 VLPs。前 S1 配体暴露在 VLPs 的表面，并显示将荧光素分子递送到肝细胞中。
新建立的掺入方法可用于开发嵌合 VLPs，通过用不同的细胞特异性配体替代前 S1 配体，这些嵌合 VLPs 可作为潜在的纳米载体，特异性地靶向各种细胞。
