The baculovirus expression vector system is a very powerful tool to produce virus-like particles and gene-therapy vectors, but the removal of co-expressed baculovirus has been a major barrier for wider industrial use. We used chimeric HIV-1 gag influenza-HA VLPs produced in Tnms42 insect cells using the baculovirus insect cell expression vector system as model VLPs. A fast and simple purification method for these VLPs with direct capture and purification within one chromatography step was developed. The insect cell culture supernatant was treated with endonuclease and filtered, before it was directly loaded onto a polymer grafted anion exchanger and eluted by a linear salt gradient. A 4.3 log clearance of baculovirus from VLPs was achieved. The absence of the baculovirus capsid protein (vp39) in the product fraction was additionally shown by HPLC-MS. When considering a vaccination dose of 10 particles, 4200 doses can be purified per L pre-treated supernatant, meeting the requirements for vaccines with <10 ng dsDNA/dose and 3.4 μg protein/dose in a single step. The process is simple with a very low number of handling steps and has the characteristics to become a platform for purification of these types of VLPs. This article is protected by copyright. All rights reserved.
This article is protected by copyright. All rights reserved.