Selective removal of DNA from protein solution with copolymer particles derived from N,N-dimethylaminopropylacrylamide.
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%.
Sakata
Masayo
M
Department of Applied Chemistry and Biochemistry, Faculty of Engineering, Kumamoto University, 2-39-1 Kurokami, Kumamoto 860-8555, Japan. msakata@gpo.kumamoto-u.ac.jp
Nakayama
Minoru
M
Kamada
Takashi
T
Kunitake
Masashi
M
Mizokami
Hiroshi
H
Hirayama
Chuichi
C
D000178:Acrylamides / Q000737:chemistry*; D000327:Adsorption; D004247:DNA / Q000302:isolation & purification*; D011506:Proteins / Q000737:chemistry*; D012996:Solutions
D000178:Acrylamides; C493602:N,N-dimethylaminopropylacrylamide; D011506:Proteins; D012996:Solutions; D004247:DNA