Bergström M, Lundblad A, Påhlsson P, Ohlson S
Department of Natural Sciences, University of Kalmar, Sweden.
J Mol Recognit. 1998 Winter;11(1-6):110-3. doi: 10.1002/(SICI)1099-1352(199812)11:1/6<110::AID-JMR402>3.0.CO;2-E.
When using weakly interacting ligands in affinity chromatography, it is possible to take advantage of a true chromatographic process in the separation, as compared with traditional affinity chromatography which is rather an on/off process. In this work, weak monoclonal antibodies were immobilized on a silica and a perfusion-type support (POROS AL) and used for high-performance liquid affinity chromatography (HPLAC). Similar carbohydrate antigens were separated under isocratic conditions according to their weak interaction with the immobilized monoclonal antibody. The affinity of the antibodies was adjusted with temperature and pH to modify the separation. The productivity of the chromatographic system was increased with the immobilized perfusion support but at the expense of loss of plate numbers. This study clearly demonstrates the potential of weak affinity biological interactions as a basis for chromatographic separation.
在亲和色谱中使用弱相互作用配体时，与传统的亲和色谱（其更像是一种开启/关闭过程）相比，有可能在分离过程中利用真正的色谱过程。在这项工作中，将弱单克隆抗体固定在硅胶和灌注型载体（POROS AL）上，并用于高效液相亲和色谱（HPLAC）。根据相似碳水化合物抗原与固定化单克隆抗体的弱相互作用，在等度条件下对其进行分离。通过调节温度和pH来调整抗体的亲和力，以改变分离效果。固定化灌注载体提高了色谱系统的生产率，但以塔板数的损失为代价。这项研究清楚地证明了弱亲和力生物相互作用作为色谱分离基础的潜力。
