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

A. Hanora, Irina Savina, F.M. Plieva, V.A. Izumrudov, B. Mattiasson, I.Y. Galaev

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 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.
Original languageEnglish
Pages (from-to)343-355
Number of pages13
JournalJournal of Biotechnology
Volume123
Issue number3
DOIs
Publication statusPublished - 29 May 2006

Fingerprint

Plasmids
DNA
Escherichia coli
Pore size
Polymers
RNA
poly(2-(dimethylamino)ethyl methacrylate)
polycations
Broncho-Vaxom
Proteins
2-(trimethylammonio)ethyl methacrylate
polyacrylamide gels

Cite this

Hanora, A. ; Savina, Irina ; Plieva, F.M. ; Izumrudov, V.A. ; Mattiasson, B. ; Galaev, I.Y. / Direct capture of plasmid DNA from non-clarified bacterial lysate using polycation-grafted monoliths. In: Journal of Biotechnology. 2006 ; Vol. 123, No. 3. pp. 343-355.
@article{abeca605aa2d40be852b4699768f5024,
title = "Direct capture of plasmid DNA from non-clarified bacterial lysate using polycation-grafted monoliths",
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 1 M NaCl as particulate-free preparation with significantly reduced content of protein and RNA as compared to the applied lysate.",
author = "A. Hanora and Irina Savina and F.M. Plieva and V.A. Izumrudov and B. Mattiasson and I.Y. Galaev",
year = "2006",
month = "5",
day = "29",
doi = "10.1016/j.jbiotec.2005.11.017",
language = "English",
volume = "123",
pages = "343--355",
journal = "Journal of Biotechnology",
issn = "0168-1656",
number = "3",

}

Direct capture of plasmid DNA from non-clarified bacterial lysate using polycation-grafted monoliths. / Hanora, A.; Savina, Irina; Plieva, F.M.; Izumrudov, V.A.; Mattiasson, B.; Galaev, I.Y.

In: Journal of Biotechnology, Vol. 123, No. 3, 29.05.2006, p. 343-355.

Research output: Contribution to journalArticle

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

VL - 123

SP - 343

EP - 355

JO - Journal of Biotechnology

JF - Journal of Biotechnology

SN - 0168-1656

IS - 3

ER -