Sakata Masayo, Nakayama Minoru, Kamada Takashi, Kunitake Masashi, Mizokami Hiroshi, Hirayama Chuichi
Department of Applied Chemistry and Biochemistry, Faculty of Engineering, Kumamoto University, 2-39-1 Kurokami, Kumamoto 860-8555, Japan.
J Chromatogr A. 2004 Mar 19;1030(1-2):117-22. doi: 10.1016/j.chroma.2003.08.109.
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%.
为了从作为药物的细胞产物中去除核酸，制备了带有阳离子官能团的交联N，N - 二甲基氨基丙基丙烯酰胺（DMP）颗粒。通过分别改变交联剂和交联中DMP的比例，可以轻松调节颗粒的疏水性及其阴离子交换容量。当使用二乙烯基苯（DVB）作为交联剂且将交联中DMP的比例调整为90摩尔％时，颗粒（DMP - DVB，90:10）对DNA（鲑鱼精巢）表现出最高的吸附活性。其吸附容量为54毫克/毫升吸附剂。另一方面，牛血清白蛋白（BSA）对DMP - DVB的吸附随着吸附剂孔径（分子量排阻；M（lim））从2×10³增加到1×10⁴而极大增加，但随着缓冲液离子强度（μ）增加到0.2或更高而降低。结果，当使用M（lim）为2×10³的DMP - DVB（80:20）作为吸附剂，在pH 7.2和μ = 0.17的条件下通过柱法时，它仅能从包含1000微克/毫升BSA和10微克/毫升DNA的BSA溶液中选择性地去除DNA。该吸附剂将BSA溶液中的DNA浓度降低至<10纳克/毫升，且BSA的回收率超过98％。
