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Selective removal of DNA from protein solution with copolymer particles derived from N,N-dimethylaminopropylacrylamide.

Authors :
Sakata M
Nakayama M
Kamada T
Kunitake M
Mizokami H
Hirayama C
Source :
Journal of chromatography. A [J Chromatogr A] 2004 Mar 19; Vol. 1030 (1-2), pp. 117-22.
Publication Year :
2004

Abstract

To remove nucleic acids from cellular products as drugs, cross-linked N,N-dimethylaminopropylacrylamide (DMP) particles with cationic functional groups were prepared. The particle's hydrophobicity and its anion-exchange capacity were easily adjusted by changing the cross-linking agent and the DMP ratio in the cross-linking, respectively. When divinylbenzene (DVB) was used as a cross-linking agent and the DMP ratio (in the cross-linking) was adjusted to 90 mol%, the particles (DMP-DVB, 90:10) showed the highest adsorbing activity of DNA (salmon spermary). Its adsorption capacity was 54 mg/ml adsorbent. On the other hand, the adsorption of bovine serum albumin (BSA) to the DMP-DVB extremely increased with increase in the adsorbent's pore size (molecular mass exclusions; M(lim)) from 2 x 10(3) to 1 x 10(4), but decreased with increase in the buffer's ionic strength (mu) to 0.2 or stronger. As a result, when the DMP-DVB (80:20) with M(lim) 2 x 10(3) was used as adsorbent by a column method at pH, 7.2 and mu = 0.17, it only selectively removed DNA from a BSA solution, including 1000 microg/ml of BSA and 10 microg/ml of DNA. The adsorbent decreased the concentration of DNA in the BSA solution to < 10 ng/ml, and the recovery rate of BSA was more 98%.

Details

Language :
English
ISSN :
0021-9673
Volume :
1030
Issue :
1-2
Database :
MEDLINE
Journal :
Journal of chromatography. A
Publication Type :
Academic Journal
Accession number :
15043261
Full Text :
https://doi.org/10.1016/j.chroma.2003.08.109