Virus-like particles (VLPs) can be used as antiviral vaccines as they mimic the structure of virus particles, with preserved conformation and immunogenicity characteristics. L1, the major capsid protein of papillomaviruses (PV) can self-assemble into VLPs currently used as highly effective vaccines. VLPs can be produced in heterologous systems, including plants. Here, a method for the expression of the L1 protein of human papillomavirus 16 (HPV 16) and the production of highly purified preparations of HPV 16 L1 VLPs is described. The method relies on the transient expression of HPV 16 L1 in Nicotiana benthamiana plants using a nonreplicating vector and on the purification of VLPs by different centrifugation steps followed by a cesium sulfate gradient. Such a procedure has also been successfully applied to other HPVs and to bovine papillomavirus 1.
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