TY - JOUR
T1 - Direct capture of plasmid DNA from non-clarified bacterial lysate using polycation-grafted monoliths
AU - Hanora, A.
AU - Savina, Irina
AU - Plieva, F.M.
AU - Izumrudov, V.A.
AU - Mattiasson, B.
AU - Galaev, I.Y.
PY - 2006/5/29
Y1 - 2006/5/29
N2 - 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 1 M NaCl as particulate-free preparation with significantly reduced content of protein and RNA as compared to the applied lysate.
AB - 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 1 M NaCl as particulate-free preparation with significantly reduced content of protein and RNA as compared to the applied lysate.
U2 - 10.1016/j.jbiotec.2005.11.017
DO - 10.1016/j.jbiotec.2005.11.017
M3 - Article
SN - 0168-1656
VL - 123
SP - 343
EP - 355
JO - Journal of Biotechnology
JF - Journal of Biotechnology
IS - 3
ER -