Details
| Original language | English |
|---|---|
| Article number | 59 |
| Journal | Microbial cell factories |
| Volume | 15 |
| Issue number | 1 |
| Publication status | Published - 8 Apr 2016 |
Abstract
Background: Production of recombinant drugs in process-friendly endotoxin-free bacterial factories targets to a lessened complexity of the purification process combined with minimized biological hazards during product application. The development of nanostructured recombinant materials in innovative nanomedical activities expands such a need beyond plain functional polypeptides to complex protein assemblies. While Escherichia coli has been recently modified for the production of endotoxin-free proteins, no data has been so far recorded regarding how the system performs in the fabrication of smart nanostructured materials. Results: We have here explored the nanoarchitecture and in vitro and in vivo functionalities of CXCR4-targeted, self-assembling protein nanoparticles intended for intracellular delivery of drugs and imaging agents in colorectal cancer. Interestingly, endotoxin-free materials exhibit a distinguishable architecture and altered size and target cell penetrability than counterparts produced in conventional E. coli strains. These variant nanoparticles show an eventual proper biodistribution and highly specific and exclusive accumulation in tumor upon administration in colorectal cancer mice models, indicating a convenient display and function of the tumor homing peptides and high particle stability under physiological conditions. Discussion: The observations made here support the emerging endotoxin-free E. coli system as a robust protein material producer but are also indicative of a particular conformational status and organization of either building blocks or oligomers. This appears to be promoted by multifactorial stress-inducing conditions upon engineering of the E. coli cell envelope, which impacts on the protein quality control of the cell factory.
Keywords
- Biodistribution, Biomaterials, E. coli, Endotoxin-free strains, Nanomedicine, Nanoparticles, Protein engineering, Recombinant proteins
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biotechnology
- Chemical Engineering(all)
- Bioengineering
- Immunology and Microbiology(all)
- Applied Microbiology and Biotechnology
Sustainable Development Goals
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In: Microbial cell factories, Vol. 15, No. 1, 59, 08.04.2016.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Structural and functional features of self-assembling protein nanoparticles produced in endotoxin-free Escherichia coli
AU - Rueda, Fabián
AU - Céspedes, María Virtudes
AU - Sánchez-Chardi, Alejandro
AU - Seras-Franzoso, Joaquin
AU - Pesarrodona, Mireia
AU - Ferrer-Miralles, Neus
AU - Vázquez, Esther
AU - Rinas, Ursula
AU - Unzueta, Ugutz
AU - Mamat, Uwe
AU - Mangues, Ramón
AU - García-Fruitós, Elena
AU - Villaverde, Antonio
N1 - Funding Information: Protein production has been partially performed by the ICTS “NANBIOSIS”, more specifically by the Protein Production Platform of CIBER in Bioengineer‑ ing, Biomaterials & Nanomedicine (CIBER‑BBN)/IBB, at the UAB (http://www. nanbiosis.es/unit/u1‑protein‑production‑platform‑ppp/). We are indebted to MINECO BIO2013‑41019‑P, AGAUR (2014SGR‑132) and CIBER de Bioingeniería, Biomateriales y Nanomedicina (project NANOPROTHER) to AV, Plan Estatal de I+D+I 2013‑2016 del Instituto de Salud Carlos III (FEDER co‑funding) FIS PI12/00327 to EV, Marató TV3 416/C/2013‑2030 to RM for funding our research. We thank the CIBER‑BBN Nanotoxicology Unit for fluorescent in vivo follow‑up using the IVIS equipment. The authors thank Manuel Hein and Dörte Grella (Research Center Borstel) and also Fran Cortes the Cell Culture and Citometry Units of the Servei de Cultius Cel·lulars, Producció d’Anticossos i Citometria (SCAC), Servei de Proteómica and to the Servei de Microscòpia, for technical assistance, and the Soft Materials Service (ICMAB‑CSIC/CIBER‑BBN). FR and MP acknowledge financial support from “Francisco Jose de Caldas” Scholarship program of COLCIENCIAS (Colombia) and Universitat Autònoma de Barcelona through pre‑doctoral fellowships respectively. UU received a Sara Borrell postdoctoral fellowship from ISCIII. AV received an ICREA ACA‑ DEMIA award. Strain KPM335 was kindly provided by Research Corporation Technologies, Tucson, AZ. Publisher Copyright: © 2016 Rueda et al.
PY - 2016/4/8
Y1 - 2016/4/8
N2 - Background: Production of recombinant drugs in process-friendly endotoxin-free bacterial factories targets to a lessened complexity of the purification process combined with minimized biological hazards during product application. The development of nanostructured recombinant materials in innovative nanomedical activities expands such a need beyond plain functional polypeptides to complex protein assemblies. While Escherichia coli has been recently modified for the production of endotoxin-free proteins, no data has been so far recorded regarding how the system performs in the fabrication of smart nanostructured materials. Results: We have here explored the nanoarchitecture and in vitro and in vivo functionalities of CXCR4-targeted, self-assembling protein nanoparticles intended for intracellular delivery of drugs and imaging agents in colorectal cancer. Interestingly, endotoxin-free materials exhibit a distinguishable architecture and altered size and target cell penetrability than counterparts produced in conventional E. coli strains. These variant nanoparticles show an eventual proper biodistribution and highly specific and exclusive accumulation in tumor upon administration in colorectal cancer mice models, indicating a convenient display and function of the tumor homing peptides and high particle stability under physiological conditions. Discussion: The observations made here support the emerging endotoxin-free E. coli system as a robust protein material producer but are also indicative of a particular conformational status and organization of either building blocks or oligomers. This appears to be promoted by multifactorial stress-inducing conditions upon engineering of the E. coli cell envelope, which impacts on the protein quality control of the cell factory.
AB - Background: Production of recombinant drugs in process-friendly endotoxin-free bacterial factories targets to a lessened complexity of the purification process combined with minimized biological hazards during product application. The development of nanostructured recombinant materials in innovative nanomedical activities expands such a need beyond plain functional polypeptides to complex protein assemblies. While Escherichia coli has been recently modified for the production of endotoxin-free proteins, no data has been so far recorded regarding how the system performs in the fabrication of smart nanostructured materials. Results: We have here explored the nanoarchitecture and in vitro and in vivo functionalities of CXCR4-targeted, self-assembling protein nanoparticles intended for intracellular delivery of drugs and imaging agents in colorectal cancer. Interestingly, endotoxin-free materials exhibit a distinguishable architecture and altered size and target cell penetrability than counterparts produced in conventional E. coli strains. These variant nanoparticles show an eventual proper biodistribution and highly specific and exclusive accumulation in tumor upon administration in colorectal cancer mice models, indicating a convenient display and function of the tumor homing peptides and high particle stability under physiological conditions. Discussion: The observations made here support the emerging endotoxin-free E. coli system as a robust protein material producer but are also indicative of a particular conformational status and organization of either building blocks or oligomers. This appears to be promoted by multifactorial stress-inducing conditions upon engineering of the E. coli cell envelope, which impacts on the protein quality control of the cell factory.
KW - Biodistribution
KW - Biomaterials
KW - E. coli
KW - Endotoxin-free strains
KW - Nanomedicine
KW - Nanoparticles
KW - Protein engineering
KW - Recombinant proteins
UR - http://www.scopus.com/inward/record.url?scp=84962917464&partnerID=8YFLogxK
U2 - 10.1186/s12934-016-0457-z
DO - 10.1186/s12934-016-0457-z
M3 - Article
C2 - 27059706
AN - SCOPUS:84962917464
VL - 15
JO - Microbial cell factories
JF - Microbial cell factories
SN - 1475-2859
IS - 1
M1 - 59
ER -