Direct capture of plasmid DNA from non-clarified bacterial lysate using polycation-grafted monoliths

Research output: Contribution to journalArticle

Abstract

Monolith columns from macroporous polyacrylamide gel were grafted with polycations, poly(N,N-dimethylaminoethyl methacrylate) (polyDMAEMA), (2-(methacryloyloxy)ethyl)-trimethyl ammonium chloride (polyMETA) and partially quaternized polyDMAEMA prepared via treating polyDMAEMA-grafted columns with propylbromide. The polymer grafting degrees varied between 34 and 110%. The polycation-grafted monolithic columns are able to capture plasmid DNA directly from alkaline lysate of Escherichia coli cells. Due to the large pore size in macroporous monoliths the particulate material present in nonclarified feeds did not block the columns. The captured plasmid DNA was eluted with I M NaCl as particulate-free preparation with significantly reduced content of protein and RNA as compared to the applied lysate. (c) 2005 Elsevier B.V. All rights reserved.

Details

Authors
  • Amro Hanora
  • Irina Savina
  • FM Plieva
  • VA Izumrudov
  • Bo Mattiasson
  • Igor Galaev
Organisations
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Industrial Biotechnology

Keywords

  • tentacle, plasmid DNA, supermacroporous gel, monolith column, chromatography
Original languageEnglish
Pages (from-to)343-355
JournalJournal of Biotechnology
Volume123
Issue number3
Publication statusPublished - 2006
Publication categoryResearch
Peer-reviewedYes