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Milligram scale par...
Milligram scale parallel purification of plasmid DNA using anion-exchange membrane capsules and a multi-channel peristaltic pump
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Syren, Per-Olof (författare)
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Rozkov, Aleksei (författare)
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Schmidt, Stefan R. (författare)
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Stroemberg, Patrik (författare)
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- Elsevier B.V. 2007
- 2007
- Engelska.
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Ingår i: Journal of chromatography. B. - : Elsevier B.V.. - 1570-0232 .- 1873-376X. ; 856, s. 68-74
- Relaterad länk:
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https://urn.kb.se/re...
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https://doi.org/10.1...
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Abstract
Ämnesord
Stäng
- A parallel chromatog. procedure for the purifn. of milligram amts. of plasmid DNA was developed. Initial studies showed that ion-exchange membrane capsules displayed high capacity for plasmid DNA. Interestingly, a weak anion exchanger (DEAE) proved to be superior to the strong quaternary ammonium group with respect to elution and regeneration properties and the 75 cm2 Sartobind D membrane capsule (MA75D, Sartorius) was selected for further studies. A method for reducing endotoxin levels by using CTAB as a precipitant was optimized. By introducing this step into the protocol, endotoxin levels could be reduced approx. 100-fold to ≤5 EU/mg plasmid. The parallel procedure was set up on a multi-channel peristaltic pump and evaluated with four different vectors (2.7-11.5 kbp). Starting with 5-10 g of E. coli cell paste (wet wt.) generally satd. the membrane adsorber, resulting in plasmid DNA yields close to 10 mg. [on SciFinder(R)]
Ämnesord
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
Nyckelord
- plasmid DNA anion exchange membrane capsule multichannel peristaltic pump
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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