A method for rapid production of heteromultimeric protein complexes in plants: assembly of protective bluetongue virus‐like particles

Summary Plant expression systems based on nonreplicating virus‐based vectors can be used for the simultaneous expression of multiple genes within the same cell. They therefore have great potential for the production of heteromultimeric protein complexes. This work describes the efficient plant‐based production and assembly of B luetongue virus‐like particles ( VLP s), requiring the simultaneous expression of four distinct proteins in varying amounts. Such particles have the potential to serve as a safe and effective vaccine against B luetongue virus ( BTV ), which causes high mortality rates in ruminants and thus has a severe effect on the livestock trade. Here, VLP s produced and assembled in N icotiana benthamiana using the cowpea mosaic virus–based H yper T rans ( CPMV ‐ HT ) and associated pEAQ plant transient expression vector system were shown to elicit a strong antibody response in sheep. Furthermore, they provided protective immunity against a challenge with a S outh A frican BTV ‐8 field isolate. The results show that transient expression can be used to produce immunologically relevant complex heteromultimeric structures in plants in a matter of days. The results have implications beyond the realm of veterinary vaccines and could be applied to the production of VLP s for human use or the coexpression of multiple enzymes for the manipulation of metabolic pathways.