Exploitation of a monoclonal antibody for weak affinity‐based separation in capillary gel electrophoresis
Exploitation of a monoclonal antibody for weak affinity‐based separation in capillary gel electrophoresis
复制标题
利用单克隆抗体进行毛细管凝胶电泳中基于弱亲和力的分离
作者：
H. Ljungberg;S. Ohlson;S. Nilsson
建立了弱生物特异性识别的高效液相色谱亲和分离方法。弱亲和层析（WAC）的使用以前受到分离效率不足的限制，仅允许获得约1000板/m。然而，研究表明，与传统色谱方法相比，用毛细管亲和凝胶电泳（CAGE）进行手性药物分离的效率大大提高。我们目前的研究证明了弱亲和单克隆抗体作为毛细管电泳免疫学分离的通用方法的潜力。单克隆抗体在硅胶毛细管内聚合，用于分离结构相似的碳水化合物抗原。结果表明，弱生物特异性相互作用可以在CAGE格式中用于对硝基苯基标记麦芽糖的α -和β -形式的高度选择性分离。然而，与亲和高效液相色谱相比，CAGE的弱亲和分离效率如何还有待观察。详细介绍了典型的分离和制备抗体凝胶。
Weak biospecific recognition has been established for affinity separation in high performance liquid chromatography (HPLC). The use of weak affinity chromatography (WAC) has been limited previously by the insufficient separation efficiency achieved, allowing only some 1000 plates/m to be obtained. However, it has been shown that chiral drug separation can be performed with capillary affinity gel electrophoresis (CAGE) at considerably improved efficiency as compared with traditional chromatographic procedures. Our present study demonstrates the potential of weak affinity monoclonal antibodies as a generic method for immunologically based separations in capillary electrophoresis. Monoclonal antibodies were polymerized within a silica capillary and were used for the separation of structurally similar carbohydrate antigens. The results indicate that weak biospecific interactions can be utilized in a CAGE format to produce highly selective separation of the α‐ and β‐forms of p‐nitro‐phenyl‐labeled maltose. It remains to be seen, however, how efficient weak affinity separation in CAGE can be compared with affinity HPLC protocols. Details of typical separations and of the preparation of the antibody gel are presented.