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

J Biotechnol. 2006 May 29;123(3):343-55. doi: 10.1016/j.jbiotec.2005.11.017. Epub 2006 Jan 10.

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 non-clarified feeds did not block the columns. The captured plasmid DNA was eluted with 1M NaCl as particulate-free preparation with significantly reduced content of protein and RNA as compared to the applied lysate.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biocompatible Materials / chemistry
  • Cell Fractionation / methods*
  • DNA, Bacterial / isolation & purification*
  • Escherichia coli / genetics*
  • Escherichia coli / isolation & purification*
  • Materials Testing
  • Membranes, Artificial*
  • Plasmids / isolation & purification*
  • Polyamines / chemistry*
  • Polyelectrolytes
  • Porosity
  • Ultrafiltration / instrumentation*
  • Ultrafiltration / methods*

Substances

  • Biocompatible Materials
  • DNA, Bacterial
  • Membranes, Artificial
  • Polyamines
  • Polyelectrolytes
  • polycations