Ljungberg H, Ohlson S, Nilsson S
Technical Analytical Chemistry, Center for Chemistry and Chemical Engineering, Lund University, Sweden.
Electrophoresis. 1998 Mar;19(3):461-4. doi: 10.1002/elps.1150190317.
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 alpha- and beta-forms of p-nitrophenyl-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.
在高效液相色谱（HPLC）中，已建立了用于亲和分离的弱生物特异性识别。以前，弱亲和色谱（WAC）的使用受到分离效率不足的限制，只能获得约1000板/米的分离效率。然而，研究表明，与传统色谱方法相比，毛细管亲和凝胶电泳（CAGE）可实现更高效率的手性药物分离。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中基于免疫分离的通用方法的潜力。单克隆抗体在硅胶毛细管内聚合，并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用于CAGE形式，以实现对对硝基苯基标记麦芽糖的α-和β-形式的高选择性分离。然而，CAGE中的弱亲和分离与亲和HPLC方案相比的效率如何，仍有待观察。本文介绍了典型分离的细节以及抗体凝胶的制备方法。
