Direct capture of plasmid DNA from non-clarified bacterial lysate using polycation-grafted monoliths.
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.
Hanora
Amro
A
Department of Biotechnology, Center for Chemistry and Chemical Engineering, Lund University, P.O. Box 124, SE-22100 Lund, Sweden.
Savina
Irina
I
Plieva
Fatima M
FM
Izumrudov
Vladimir A
VA
Mattiasson
Bo
B
Galaev
Igor Yu
IY
D001672:Biocompatible Materials / Q000737:chemistry; D002458:Cell Fractionation / Q000379:methods*; D004269:DNA, Bacterial / Q000302:isolation & purification*; D004926:Escherichia coli / Q000235:genetics* / Q000302:isolation & purification*; D008422:Materials Testing; D008567:Membranes, Artificial; D010957:Plasmids / Q000302:isolation & purification*; D011073:Polyamines / Q000737:chemistry*; D000071228:Polyelectrolytes; D016062:Porosity; D014462:Ultrafiltration / Q000295:instrumentation* / Q000379:methods*
D001672:Biocompatible Materials; D004269:DNA, Bacterial; D008567:Membranes, Artificial; D011073:Polyamines; D000071228:Polyelectrolytes