Open AccessAll-in-one biofabrication and loading of recombinant vaults in human cells
Author(s) -
Fernando Martín,
Aida Carreño,
Rosa Mendoza,
Pablo Caruana,
Francisco Rodríguez,
Marlon Bravo,
Antoni Benito,
Neus Ferrer-Miralles,
María Virtudes Céspedes,
José Luís Corchero
Publication year - 2022
Publication title -
biofabrication
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.328
H-Index - 80
eISSN - 1758-5090
pISSN - 1758-5082
DOI - 10.1088/1758-5090/ac584d
Subject(s) - biofabrication , recombinant dna , synthetic biology , computational biology , computer science , nanotechnology , tissue engineering , chemistry , biology , materials science , gene , biomedical engineering , engineering , biochemistry
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.