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Milligram scale parallel purification of plasmid DNA using anion-exchange membrane capsules and a multi-channel peristaltic pump
2007 (English)In: Journal of chromatography. B, ISSN 1570-0232, E-ISSN 1873-376X, Vol. 856, 68-74 p.Article in journal (Refereed) Published
Abstract [en]

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 ‚ȧ5 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)]

Place, publisher, year, edition, pages
Elsevier B.V. , 2007. Vol. 856, 68-74 p.
Keyword [en]
plasmid DNA anion exchange membrane capsule multichannel peristaltic pump
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:kth:diva-185577ISI: 000249615100011PubMedID: 17604704Scopus ID: 2-s2.0-34548182829OAI: oai:DiVA.org:kth-185577DiVA: diva2:922313
Note

QC 20160422

Available from: 2016-04-22 Created: 2016-04-22 Last updated: 2016-04-22Bibliographically approved

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Syren, Per-Olof
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