| 摘要 |
<p>Methods for obtaining HCV complexes and HCV-like particles comprising HCV structural genes are provided. In one method, cells containing HCV-like particles are lysed with digitonin in the presence of protease inhibitors. Polyethylene glycol is slowly added to the lysate, to provide a precipitate that comprises complexes of the HCV structural proteins associated with lipi d vesicles or micelles and complexes comprising viral structural proteins in t he form of insoluble aggregates. In another method, the lysate is centrifuged through a sucrose cushion. Preferably, the pellet is then subjected to equilibrium ultracentrifugation, to provide a preparation of HCV-like particles that are heterogenous in size. The third method comprises subjecti ng the infected cells to hypertonic/hypotonic shock, and lysing the cells with digitonin in the presence of protease inhibitors. The lysate is pelleted and fractionated to provide a population of HCV-like particles that are substantially homogenous and have an average diameter of about 50 nm. As use d herein the term " substantially homogenous" means that the shape of the particles are similar and that the size of the particles vary by 10% or less . Methods of using the HCV complexes and HCV-like particles as screening tools , diagnostic tools, and immunogeniccompositions are also provided. Methods of treating patients exhibiting symptoms of HCV infection with compounds or substances that interfere with binding or internalization of the present HCV - like particles to asialoglycoprotein receptors are also provided.</p> |
