Abstract
Baculovirus is capable of transducing mammalian cells with high efficiency and has been proved to be a promising vector for in vivo or ex vivo gene therapy. In the context of vaccine and gene therapy applications, high purity of baculovirus is necessary, thus we aimed at developing a simple and rapid purification process. In this study, we used a recombinant baculovirus with Vesicular Stomatitis Virus Glycoprotein（VSV-G）on the viral surface, which has been shown to enhance viral transduction efficiency. We also demonstrated that the pseudotyped viral vector can improve viral stability. In order to develop a simple and rapid purification process for recombinant baculovirus, we used tangential flow filtration（TFF）and Concanavalin A（Con A）affinity Chromatography in series for high purity purification. The former is a convenient method to remove host cell proteins and concentrate recombinant baculovirus prior to chromatographic step with a higher recovery of 75%, and the latter leads to approximately 21% recovery of recombinant baculovirus by elution with methyl-alpha-D-mannopyranoside. The newly developed method resulted in an overall recovery yield（~15%）that was higher than those resulting from immobilized metal affinity chromatography（1~2%）and gradient ultracentrifugation（<1%）. In addition, rapid and large-scale purification and high recovery yield of baculovirus can be achieved using Con A affinity chromatography. This is the first report describing the method of TFF combined with Con A affinity chromatography for baculovirus purification.