Titlebar

Bibliografische Daten exportieren
Literatur vom gleichen Autor
plus auf ERef Bayreuth
plus bei Google Scholar

 

Proposal for a better integration of bacterial lysis into the production of plasmid DNA at large scale.

Titelangaben

O'Mahony, Kevin ; Freitag, Ruth ; Hilbrig, Frank ; Müller, Patrick ; Schumacher, Ivo:
Proposal for a better integration of bacterial lysis into the production of plasmid DNA at large scale.
In: Journal of Biotechnology. Bd. 119 (September 2005) Heft 2 . - S. 118-132.
ISSN 1873-4863
DOI: https://doi.org/10.1016/j.jbiotec.2005.03.020

Volltext

Link zum Volltext (externe URL): Volltext

Abstract

The paper addresses the question of how to achieve bacterial lysis in large-scale plasmid DNA production processes, where conventional alkaline lysis may become awkward to handle. Bacteria were grown in shaker flasks and a bioreactor. Suboptimal growth conditions were found advantageous for stable plasmid production at high copy numbers (up to 25mg/L could be achieved). Cells were harvested by filtration in the presence of a filter aid. A linear relationship between the biomass and the optimal filter aid concentration in terms of back pressure could be established. Bacteria-containing filter cakes were washed with isotonic buffer and lysis was achieved in situ by a two-step protocol calling for fragilisation of the cells followed by heat lysis in a suitable buffer. RNA and other soluble cell components where washed out of the cake during this step, while the plasmid DNA was retained. Afterwards a clear lysate containing relatively pure plasmid DNA could be eluted from the cake mostly as the desired supercoiled topoisomer, while cell debris and genomic DNA were retained. Lysis is, thus, integrated not only with cell capture but also with a significant degree of isolation/purification, as most impurities were considerably reduced during the procedure.

Weitere Angaben

Publikationsform: Artikel in einer Zeitschrift
Begutachteter Beitrag: Ja
Institutionen der Universität: Fakultäten > Fakultät für Ingenieurwissenschaften > Lehrstuhl Bioprozesstechnik > Lehrstuhl Bioprozesstechnik - Univ.-Prof. Dr. Ruth Freitag
Fakultäten
Fakultäten > Fakultät für Ingenieurwissenschaften
Fakultäten > Fakultät für Ingenieurwissenschaften > Lehrstuhl Bioprozesstechnik
Titel an der UBT entstanden: Nein
Themengebiete aus DDC: 500 Naturwissenschaften und Mathematik
600 Technik, Medizin, angewandte Wissenschaften
600 Technik, Medizin, angewandte Wissenschaften > 620 Ingenieurwissenschaften
Eingestellt am: 25 Feb 2016 14:23
Letzte Änderung: 25 Feb 2016 14:23
URI: https://eref.uni-bayreuth.de/id/eprint/31055