Leickt L, Grubb A, Ohlson S
Department of Natural Sciences, University of Kalmar, Sweden.
J Mol Recognit. 1998 Winter;11(1-6):114-6. doi: 10.1002/(SICI)1099-1352(199812)11:1/6<114::AID-JMR403>3.0.CO;2-#.
As the interest in weak-affinity antibodies has been widened by their introduction to various analytical techniques such as HPLC, capillary electrophoresis and biosensors, there has been a need for new screening/monitoring methods. In this study, weak-affinity chromatography was adopted to screen/monitor directly for monoclonal antibodies in ascites. Monoclonal antibodies against a carbohydrate antigen (maltohexaose) were used to evaluate this approach. In short, malthohexaose was immobilized on an HPLC support in such a configuration to allow, during HPLC, retardation of weak monoclonal antibodies. Based on the retention, the affinity or the avidity, as determined by the presence of multiple binding of the monoclonal antibody towards antigen, can be estimated. In this way it is possible to select clones of hybridomas that produce desired weak monoclonal antibodies. Adjustments in temperature (10-20 degrees C) were used to moderate the retention and hence affinity of the weak monoclonal antibodies during chromatography.
随着弱亲和力抗体通过引入各种分析技术（如高效液相色谱法、毛细管电泳法和生物传感器）而受到更广泛关注，对新的筛选/监测方法的需求应运而生。在本研究中，采用弱亲和力色谱法直接筛选/监测腹水中的单克隆抗体。使用针对碳水化合物抗原（麦芽六糖）的单克隆抗体来评估该方法。简而言之，麦芽六糖以这样一种配置固定在高效液相色谱支持物上，使得在高效液相色谱过程中，弱单克隆抗体能够被滞留。基于保留情况，可以估计由单克隆抗体与抗原的多重结合所确定的亲和力或亲合力。通过这种方式，可以选择产生所需弱单克隆抗体的杂交瘤克隆。在色谱过程中，通过调节温度（10 - 20摄氏度）来调节弱单克隆抗体的保留情况，进而调节其亲和力。
