Nanobiotechnology: “All-in-one biofabrication and loading of recombinant vaults in human cells”

Biofabrication, Volume 14, Number 2



One of the most promising approaches in the drug delivery field is the use of naturally occurring self-assembling protein nanoparticles, such as virus-like particles, bacterial microcompartments or vault ribonucleoprotein particles as drug delivery systems (DDSs). Among them, eukaryotic vaults show a promising future due to their structural features, in vitro stability and non-immunogenicity. Recombinant vaults are routinely produced in insect cells and purified through several ultracentrifugations, both tedious and time-consuming processes. As an alternative, this work proposes a new approach and protocols for the production of recombinant vaults in human cells by transient gene expression of a His-tagged version of the major vault protein (MVP-H6), the development of new affinity-based purification processes for such recombinant vaults, and the all-in-one biofabrication and encapsulation of a cargo recombinant protein within such vaults by their co-expression in human cells. Protocols proposed here allow the easy and straightforward biofabrication and purification of engineered vaults loaded with virtually any INT-tagged cargo protein, in very short times, paving the way to faster and easier engineering and production of better and more efficient DDS.

Fernando Martín7,1, Aida Carreño8,1, Rosa Mendoza1,2, Pablo Caruana3, Francisco Rodriguez3, Marlon Bravo4,5, Antoni Benito4,5, Neus Ferrer-Miralles1,2,6, M Virtudes Céspedes9,3 and José Luis Corchero9,1,2,6