In Vitro Disassembly and Reassembly of Triple-Layered Rotavirus-Like Particles

摘要

Virus-like particles (VLPs) are of interest in vaccination, gene therapy and drug delivery. Their assembly process needs to be understood in order to improve their yield and quality. Triple-layered Rotavirus-like particle (TLP 2/6/7), a candidate vaccine against Rotavirus infection, is produced in insect cells by co-infection of baculoviruses coding for VP2, VP6 and VP7, the main structural proteins. Our earlier results have shown that the outer layer, constituted by VP7 monomers, is unstable, as VP7 can peel off resulting in the formation of an uncoated double-layered particle 2/6 (DLP). In this work, we investigated the parameters involved in the disassembly of TLP 2/6/7 into DLP 2/6. For this, purified TLP 2/6/7 was used for in vitro disassembly. Next, DLP 2/6 was assembled into TLP 2/6/7 by the addition of purified VP7 monomers. The kinetics of such disassembly and reassembly reactions, as monitored by light scattering spectroscopy, were found to be first and second order, respectively. The reaction constants were calculated at different temperatures, reactants and ionic strengths.