6 research outputs found
Detoxifying Escherichia coli for endotoxin-free production of recombinant proteins
Es va publicar un treball amb esmenes a aquest article que es pot consultar a https://ddd.uab.cat/record/185349 (DOI 10.1186/s12934-015-0265-x)Background: lipopolysaccharide (LPS), also referred to as endotoxin, is the major constituent of the outer leaflet of the outer membrane of virtually all Gram-negative bacteria. The lipid A moiety, which anchors the LPS molecule to the outer membrane, acts as a potent agonist for Toll-like receptor 4/myeloid differentiation factor 2-mediated pro-inflammatory activity in mammals and, thus, represents the endotoxic principle of LPS. Recombinant proteins, commonly manufactured in Escherichia coli, are generally contaminated with endotoxin. Removal of bacterial endotoxin from recombinant therapeutic proteins is a challenging and expensive process that has been necessary to ensure the safety of the final product. -Results: as an alternative strategy for common endotoxin removal methods, we have developed a series of E. coli strains that are able to grow and express recombinant proteins with the endotoxin precursor lipid IVA as the only LPS-related molecule in their outer membranes. Lipid IVA does not trigger an endotoxic response in humans typical of bacterial LPS chemotypes. Hence the engineered cells themselves, and the purified proteins expressed within these cells display extremely low endotoxin levels. - Conclusions: this paper describes the preparation and characterization of endotoxin-free E. coli strains, and demonstrates the direct production of recombinant proteins with negligible endotoxin contamination
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Solid freeform fabrication of highly loaded composite materials
Composites are known for their unique blend of modulus, strength, and toughness. This study focuses on two types of composites; organic-inorganic hybrids and the mineralization of highly swollen polymer gels. Both of these composite systems mimic the biological process of composite formation, known as biomineralization. Biomineralization allows for the control of the precipitating phase through an interaction with the organic matrix. This allows higher volume fractions of inorganic material than can be achieved by many traditional processing techniques. Solid freeform fabrication is a processing method that builds materials by the sequential addition of thin layers. As long as the material can easily be converted from a liquid to a solid, it should be amenable for this processing technique. Freeform fabrication has three distinctions from traditional processing techniques that may enable the formation of composite materials with improved mechanical properties. These are the sequential addition of layers, which allows a layer by layer influence of chemistry, the ability to form complex geometries, and finally, extrusion freeform fabrication has been shown to align fibers due to the extrusion of the slurry through a needle. Cracking and shrinkage still play a major role in forming solid parts. The use of an open mesh structure in combination with proper materials selection allowed the formation of highly loaded composite materials without cracking. The modulus values of these materials ranged from 0.1 GPa to 6.0 GPa. The mechanical properties of these materials were modeled
Erratum to: Detoxifying Escherichia coli
Aquest text recull una esmena a l'article que es pot consultar a https://ddd.uab.cat/record/132806 (DOI: 10.1186/s12934-015-0241-5
Erratum to : Detoxifying Escherichia coli for endotoxin-free production of recombinant proteins
Aquest text recull una esmena a l'article que es pot consultar a https://ddd.uab.cat/record/132806 (DOI: 10.1186/s12934-015-0241-5
Detoxifying Escherichia coli for endotoxin-free production of recombinant proteins
Es va publicar un treball amb esmenes a aquest article que es pot consultar a https://ddd.uab.cat/record/185349 (DOI 10.1186/s12934-015-0265-x)Background: lipopolysaccharide (LPS), also referred to as endotoxin, is the major constituent of the outer leaflet of the outer membrane of virtually all Gram-negative bacteria. The lipid A moiety, which anchors the LPS molecule to the outer membrane, acts as a potent agonist for Toll-like receptor 4/myeloid differentiation factor 2-mediated pro-inflammatory activity in mammals and, thus, represents the endotoxic principle of LPS. Recombinant proteins, commonly manufactured in Escherichia coli, are generally contaminated with endotoxin. Removal of bacterial endotoxin from recombinant therapeutic proteins is a challenging and expensive process that has been necessary to ensure the safety of the final product. -Results: as an alternative strategy for common endotoxin removal methods, we have developed a series of E. coli strains that are able to grow and express recombinant proteins with the endotoxin precursor lipid IVA as the only LPS-related molecule in their outer membranes. Lipid IVA does not trigger an endotoxic response in humans typical of bacterial LPS chemotypes. Hence the engineered cells themselves, and the purified proteins expressed within these cells display extremely low endotoxin levels. - Conclusions: this paper describes the preparation and characterization of endotoxin-free E. coli strains, and demonstrates the direct production of recombinant proteins with negligible endotoxin contamination