A Technique for the Generation of Influenza Virus-Like Particles via a Mammalian System
Published: September 29, 2023
Abstract
Source: Arevalo, M. T., et al. Expression and Purification of Virus-like Particles for Vaccination. J. Vis. Exp. (2016).
This video demonstrates the generation of influenza virus-like particles (VLPs) in a mammalian system. Upon expressing three eukaryotic expression vectors — encoding different viral structural proteins — in suitable mammalian host cells, the expressed viral proteins self-assemble into VLPs on the host plasma membrane and release via budding from the cell surface.
Protocol
1. Mammalian Expression System for Generation of Influenza H3N2 VLP Subclone viral structural glycoproteins hemagglutinin (HA), neuraminidase (NA), and human immunodeficiency virus (HIV) Gag (group-specific antigen) p55 into eukaryotic expression vectors, such as previously described pTR600. Amplify DNA in chemically competent Escherichia coli (e.g., DH5α) and isolate transfection-grade plasmid using a plasmid purification kit per manufacturer's instructio…
Disclosures
The authors have nothing to disclose.
Materials
| Plasmid Maxi Kit | Qiagen | 12163 | |
| DMEM | Cellgro | 10-013 | |
| Lipofectamine | Life Technologies | L3000075 | Lipofectamine 2000, Lipofectamine 3000 |
| Opti-MEM I | Life Technologies | 31985062 | |
| Cimarec I 6 multipoint stirrer plate | ThermoFisher Scientific | 50094596 | |
| Polyclear ultracentrifuge tubes | Seton | 7025 | |
| 0.22 μm vacuum filter top (500 mL) | Nalgene | 569-0020 | |
| Glycerol | Sigma | G5516 |
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Cite This Article
A Technique for the Generation of Influenza Virus-Like Particles via a Mammalian System. J. Vis. Exp. (Pending Publication), e21700, doi: (2023).