title: A protocol for the gentle purification of virus-like particles produced in plants.
abstract: The purpose of the protocol is to extract and purify virus-like particles (VLPs) that have been produced in plants. More specifically, this method is well suited to the purification of chimaeric and genetically modified VLPs that do not have native surface properties. This will be the case for VLPs used in antigen display experiments. Such particles are often more fragile than their wild-type infectious virus counterparts, and as such can be damaged or lost during procedures that involve pelleting or precipitating the particles. The method presented here is based on ultracentrifugation and density gradients, with no pelleting or precipitation step. It makes virtually no assumptions about the yield of recombinant VLPs or their properties, which means that this protocol is ideally suited to screening new constructs which are expected to lead to the formation of VLPs. This protocol will allow the researcher to determine whether the construct does indeed form VLPs, and if it does, will reduce the likelihood of those particles being lost or damaged during the purification process. Because of its non-specific nature, this protocol may also be suited to the purification of viruses of unknown nature from leaf material where an infection is suspected.
authors.affiliation: Department of Biological Chemistry, John Innes Centre, Norwich, UK. Electronic address: Hadrien.peyret@jic.ac.uk.
authors.forename: Hadrien
authors.initials: H
authors.lastname: Peyret
mesh_terms: D010944:Plants / Q000235:genetics / Q000378:metabolism*; D014461:Ultracentrifugation / Q000379:methods*; D014773:Virology / Q000379:methods*; D022701:Virosomes / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D022701:Virosomes
keywords: Molecular farming; Nycodenz gradient; Purification; Sucrose cushion; VLP; Virus-like particle