Home > Articles > Published articles > All-in-one biofabrication and loading of recombinant vaults in human cells |
Date: | 2022 |
Abstract: | 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. |
Grants: | Instituto de Salud Carlos III PI17/00553 Instituto de Salud Carlos III PI17/00150 Instituto de Salud Carlos III PI20/00770 Instituto de Salud Carlos III PI20/00623 |
Note: | Altres ajuts: Fundación Mutua Madrileña (FMMA) through project 'Targeted therapy for selective elimination of metastatic stem cells CXCR4+ in endometrial cancer' (AP1666942017) |
Note: | Altres ajuts: Asociación Española Contra el Cancer (AECC) through project 'Development of an antitumor protein delivery system into ovarian cancer cells using the subcellular vault' (IDEAS18038BENI) |
Rights: | Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original. |
Language: | Anglès |
Document: | Article ; recerca ; Versió publicada |
Subject: | Vault particles ; Protein nanocages ; Drug delivery systems ; Self-assembling protein nanoparticles ; Eukaryotic cell factories |
Published in: | Biofabrication, Vol. 14, Issue 2 (April 2022) , art. 025018, ISSN 1758-5090 |
17 p, 2.2 MB |