The incorporation of the A2 protein to produce novel Qβ virus‐like particles using cell‐free protein synthesis

Virus‐like particles (VLPs) have been employed for a number of nanometric applications because they self‐assemble, exhibit a high degree of symmetry, and can be genetically and chemically modified. However, high symmetry does not allow for a single unique modification site on the VLP. Here, we demonstrate the co‐expression of the cytotoxic A2 protein and the coat protein of the bacteriophage Qβ to form a nearly monodispersed population of novel VLPs. Cell‐free protein synthesis allows for direct access and optimization of protein‐synthesis and VLP‐assembly. The A2 is shown to be incorporated at high efficiency, approaching a theoretical maximum of one A2 per VLP. This work demonstrates de novo production of a novel VLP, which contains a unique site that has the potential for future nanometric engineering applications. © 2011 American Institute of Chemical Engineers Biotechnol. Prog., 2012