Isolation of cell-free bacterial inclusion bodies.
BACKGROUND
Bacterial inclusion bodies are submicron protein clusters usually found in recombinant bacteria that have been traditionally considered as undesirable products from protein production processes. However, being fully biocompatible, they have been recently characterized as nanoparticulate inert materials useful as scaffolds for tissue engineering, with potentially wider applicability in biomedicine and material sciences. Current protocols for inclusion body isolation from Escherichia coli usually offer between 95 to 99% of protein recovery, what in practical terms, might imply extensive bacterial cell contamination, not compatible with the use of inclusion bodies in biological interfaces.


RESULTS
Using an appropriate combination of chemical and mechanical cell disruption methods we have established a convenient procedure for the recovery of bacterial inclusion bodies with undetectable levels of viable cell contamination, below 10⁻¹ cfu/ml, keeping the particulate organization of these aggregates regarding size and protein folding features.


CONCLUSIONS
The application of the developed protocol allows obtaining bacterial free inclusion bodies suitable for use in mammalian cell cultures and other biological interfaces.
Rodríguez-Carmona
Escarlata
E
Institut de Biotecnologia i de Biomedicina and Departament de Genètica i de Microbiologia, Universitat Autònoma de Barcelona, 08193 Bellaterra (Cerdanyola del Vallès), Barcelona, Spain.
Cano-Garrido
Olivia
O
Seras-Franzoso
Joaquin
J
Villaverde
Antonio
A
García-Fruitós
Elena
E
D004926:Escherichia coli / Q000378:metabolism; D002479:Inclusion Bodies / Q000737:chemistry* / Q000378:metabolism; D009113:Muramidase / Q000378:metabolism; D011312:Pressure; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013010:Sonication; D013501:Surface-Active Agents / Q000737:chemistry
D011994:Recombinant Proteins; D013501:Surface-Active Agents; D009113:Muramidase