摘要
作者采用ＤＥＡＥ－４０ＨＲ阴离子交换柱。在快速蛋白液相色谱系统（ＦＰＬＣ）建立了ＩｇＧ类单克隆抗体制备级纯化方法。该法是将ＭｃＡｂ腹水经５０％饱和硫酸铵盐析粗分离后，再用阴离子交换色谱纯化，一次上样量相当于８０～１００ｍｌ腹水。可得纯化ＭｃＡｂ５００～６００ｍｇ，得率为６～７ｍｇ／ｍｌ腹水，回收率为５７～６７％．一次纯化周期仅需４５ｍｉｎ。经ＳＤＳ－ＰＡＧＥ检测ＭｃＡｂ纯度为９５％，ＡＢＣ染色法测定ＭｃＡｂ活性为１：２００００（３．１３×１０－１０ｍｏｌ／Ｌ）．
The purification of IgG McAb on preparative scale by fast protein liquid chromatography(FPLC) on an anion exchange column has been established.The antihepatoma McAb HAb18 was ptirified from ascitic fluid by precipitation with 50 %saturated ammonium sulfate,followed by,column of DEAE-HR40 with pH7.4 PB,and 500～600 mg of the purified IgG was obtained in 45 min at one operating cycle. The recovery rate of IgG was 57%～67%, with purity of McAb being about 95% on SDSPAGE. And immunoactivity was detected at 3.13×10-10 mol/L by ABC immunohistoc hemistry.
基金
国家863课题
关键词
抗体
液相色谱法
单克隆抗体
antibody, monoclonal
purification
liquid chromatography
