Clezardin P, McGregor J L, Manach M, Boukerche H, Dechavanne M
J Chromatogr. 1985 Jan 25;319(1):67-77. doi: 10.1016/s0021-9673(01)90540-0.
A one-step chromatographic procedure was used to isolate rapidly mouse IgG monoclonal antibodies (mAbs) (IgG1, IgG2a, IgG2b) contained in ascites fluids and Fab fragments contained in papain-treated mAb suspensions. Chromatographic separations were performed on an anion-exchange Mono Q column connected to a fast protein liquid chromatographic (FPLC) system. Detection of mAb or their antigen binding fragments (Fab) in eluted peaks was performed using sodium dodecyl sulphate-polyacrylamide gel electrophoresis together with a silver or a Coomassie Brillant Blue R 250 staining technique and solid phase radioimmunoassay with 125I-labelled sheep anti-mouse antibodies directed against total immunoglobulins. Rapid assessment of the purity of isolated mAbs and their Fab fragments was performed by gel permeation chromatography on a TSK G 3000 SW column. Mouse mAbs and their Fab fragments were rapidly isolated (25 min), in a functionally active state, to a high degree of purity on the FPLC-Mono Q system compared to the time taken by other techniques.
采用一步色谱法快速分离腹水液中所含的小鼠IgG单克隆抗体（mAb）（IgG1、IgG2a、IgG2b）以及木瓜蛋白酶处理的mAb悬浮液中所含的Fab片段。色谱分离在连接到快速蛋白质液相色谱（FPLC）系统的阴离子交换Mono Q柱上进行。使用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳结合银染或考马斯亮蓝R 250染色技术以及用针对总免疫球蛋白的125I标记羊抗小鼠抗体进行的固相放射免疫测定法，对洗脱峰中的mAb或其抗原结合片段（Fab）进行检测。通过在TSK G 3000 SW柱上进行凝胶渗透色谱法，对分离得到的mAb及其Fab片段的纯度进行快速评估。与其他技术所需时间相比，在FPLC-Mono Q系统上，小鼠mAb及其Fab片段能够快速（25分钟）分离出来，且处于功能活性状态，纯度很高。
