Sakata M, Sueda T, Ihara H, Hirayama C
Department of Applied Chemistry, Kumamoto University, Japan.
Chem Pharm Bull (Tokyo). 1996 Feb;44(2):328-32. doi: 10.1248/cpb.44.328.
Novel copolymeric adsorbents for the selective removal of endotoxin from an acidic protein solution were prepared. The adsorbents comprise spherical copolymers derived from N,N-dimethylaminopropylacrylamide (DMAPAA) and divinylbenzene (DVB). When the molar ratio of DMAPAA to DVB was 80/20 (amino-group content: 5.1 meq/g) and the pore size (molecular mass exclusion of polysaccharide, Mlim) was 4000 to 10000, DMAPAA/DVB showed high endotoxin-adsorbing activity at pH 5.0 to 9.0 and ionic strengths of mu = 0.05 to 0.4. The capacity of the adsorbent (Mlim: 4000) was 390 micrograms of endotoxin (lipopolysaccharide purified from E. coli O111:B4) per ml of the adsorbent using the batchwise method. The apparent dissociation constant between endotoxin and the adsorbent was 2.2 x 10(-12) M. On the other hand, the adsorption of bovine serum albumin, an acidic protein, by the adsorbent increased with an increase in Mlim from 4000 to 10000, but decreased with an increase in ionic strength (mu) from 0.05 to 0.2. As a result, DMAPAA/DVB (80/20) (Mlim: 4000) selectively removed endotoxin from various acidic protein solutions at pH 7.0 and mu = 0.05. The residual concentration of endotoxin in the protein solution always decreased to a concentration lower than 0.1 ng/ml, and recovery of the protein was more than 97%.
制备了用于从酸性蛋白质溶液中选择性去除内毒素的新型共聚吸附剂。这些吸附剂包含由N,N - 二甲基氨基丙基丙烯酰胺(DMAPAA)和二乙烯基苯(DVB)衍生的球形共聚物。当DMAPAA与DVB的摩尔比为80/20(氨基含量:5.1 meq/g)且孔径(多糖的分子量排阻,Mlim)为4000至10000时,DMAPAA/DVB在pH 5.0至9.0以及离子强度μ = 0.05至0.4的条件下表现出高内毒素吸附活性。使用分批法时,吸附剂(Mlim:4000)的容量为每毫升吸附剂390微克内毒素(从大肠杆菌O111:B4纯化的脂多糖)。内毒素与吸附剂之间的表观解离常数为2.2×10^(-12) M。另一方面,吸附剂对酸性蛋白质牛血清白蛋白的吸附随着Mlim从4000增加到10000而增加,但随着离子强度(μ)从0.05增加到0.2而降低。结果,DMAPAA/DVB(80/20)(Mlim:4000)在pH 7.0和μ = 0.05时从各种酸性蛋白质溶液中选择性地去除内毒素。蛋白质溶液中内毒素的残留浓度总是降低到低于0.1 ng/ml的浓度,并且蛋白质的回收率超过97%。