Team:UCL London/Research/Extractery
From 2011.igem.org
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- | The most difficult section of any industrial grade bio-manufacturing process is the downstream processing involved in product recovery. And this is absolutely crucial in the case of plasmid DNA manufacturing, as the DNA plasmids need to be purified to a very high standard and quality, according to the FDA guidelines, in order to minimise the incidence of side effects following clinical administration. Furthermore, plasmid DNA purification poses a harder challenge than normal heterologous protein expression due to the close resemblance in structure and property of plasmid DNA with other cellular DNAs and RNAs. Current industry processes involves bursting the cultured cells open using time-consuming heat or alkaline lysis step and then isolating the supercoiled plasmids from impurities like proteins, endotoxins, genomic DNA, RNA and other unwanted isoforms of the plasmid (linear, multimers, open circle and irreversible damaged plasmids) using extensive and expensive purifications techniques. Our extractory sub project aims to make this entire purification process much simpler, more efficient and also automated to a certain extent inside E coili. | + | <html><div style="float:right"><iframe width="560" height="315" src="http://www.youtube.com/embed/V68BQm-JZr4" frameborder="0" allowfullscreen></iframe></div></html>The most difficult section of any industrial grade bio-manufacturing process is the downstream processing involved in product recovery. And this is absolutely crucial in the case of plasmid DNA manufacturing, as the DNA plasmids need to be purified to a very high standard and quality, according to the FDA guidelines, in order to minimise the incidence of side effects following clinical administration. Furthermore, plasmid DNA purification poses a harder challenge than normal heterologous protein expression due to the close resemblance in structure and property of plasmid DNA with other cellular DNAs and RNAs. Current industry processes involves bursting the cultured cells open using time-consuming heat or alkaline lysis step and then isolating the supercoiled plasmids from impurities like proteins, endotoxins, genomic DNA, RNA and other unwanted isoforms of the plasmid (linear, multimers, open circle and irreversible damaged plasmids) using extensive and expensive purifications techniques. Our extractory sub project aims to make this entire purification process much simpler, more efficient and also automated to a certain extent inside E coili. |
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