We have a customer who wants to use baculovirus to expression virus-like particles. Does anyone have experience with this and can give some tips?
We have not done it ourselves and I have heard from others in our university who have tried that it is not so easy... But as there are publications it must be possible. We are actually considering making VLPs in CHO cells or similar. Does anyone have experience with this?
Imre Berger mentioned VLP expression in insect cells in one of his talks. I'm not sure if he did it himself or mentioned a collaborator who was doing it. Hope that helps.
We have produced a few types of VLPs (non-enveloped and enveloped) using insect cells. Actually baculovirus/insect cell expression system is a good system for producing VLPs. The yields can be high for some VLPs (>500mg/L). Depending on whether you are expressing one or multiple proteins to form the VLP, you can either co-infect the cells with multiple recombinant baculoviruses or using dual vectors to allow co-expression of 2 proteins. I haven't tried the mulitBac system. Insect cell enables the VLP to self-assemble and the assembled VLPs are purified from either the cells or supernatant. We use Asymmetric-Flow Field-Flow Fractionation and TEM for the analysis of purified VLPs.
I may be able to provide more specific tips if I know what VLPs you would like to produce. Feel free to contact me.
All the best,
I was also asked to produce VLPs. Is there any change to the protocol to obtain VLPs?
No significant changes to the expression protocol for VLPs. We do the usual, which is a timecourse study to determine optimal time of harvest, check the effect of cell lines, temp etc. If the VLP is made up of 2 proteins or more, it's important to ensure that all proteins are expressed and the assembled VLPs consist of these proteins. If I using 2 separate virus stocks to co-infect a culture, I will also check the expression of each protein separately. Sometimes, I do observe the preference expression of one protein over the other during a co-expression (MOI/TOI is kept the same).
Thanks for the comments. Linda, I will contact you with more details about the project, thanks for the offer.