Attempted synthesis of taxol utilizing anti-taxol monoclonal antibody as a as a chiral mould

制备了2‘-琥珀基紫杉醇-BSA和2’-琥珀基紫杉醇-RSA偶联物。半抗原和蛋白质之间的连接通常通过紫外法或荧光胺法来确认。然而，由于2‘-琥珀酰基紫杉醇和蛋白质的紫外吸收峰相似，用这些方法很难确定2’-琥珀酰基紫杉醇与蛋白质的结合。因此，我们使用中性损失扫描和MS/MS技术进行了质谱分析，以确认2‘-琥珀酸紫杉醇-蛋白质偶联物的配方。免疫BALB/c小鼠。间隔2周，每次2‘-琥珀基紫杉醇-RSA。小鼠的脾细胞用常规方法处理，得到四个杂交瘤细胞，产生抗紫杉醇的单抗。但杂交瘤细胞产生的抗紫杉醇单抗能力不强，抗体效价较低。不幸的是，随着克隆的进行，抗体的产量逐渐下降。尽管我们在抗体的数量、佐剂的种类以及培养条件等方面做出了努力，但这个问题并没有得到改善。初步尝试了巴卡菌素III与活化的C13-羧酸酯(维尼酯)在粗抗体作用下的缩合反应。由于抗体产生功能丧失，尚未对抗体反应进行进一步检查。

2'-Succinyltaxol-BSA and 2'-succinyltaxol-RSA conjugates were prepared. Formation of a linkage between hapten and protein is usually confirmed by the UV or fluorescamine method. However, it was difficult to confirm the binding of 2'-succinyltaxol to the protein by these methods owing to the similar UV absorption maxima of 2'-succinyltaxol and protein. We therefore conducted a mass spectrometric analysis using the neutral loss scan and MS/MS techniques to confirm the formulation of the 2'-succinyltaxol-protein conjugate. Balb/c mice were immunized i.c. at 2-week intervals each with 2'-Succinyltaxol-RSA. The spleen cells from the mice were treated in a conventional way, giving four hybridomas producing an anti-taxol monoclonal antibody. However, the hybridomas did not have a high production ability of an anti-taxol monoclonal antibody, and their antibody titers were rather low. Unfortunately, production of the antibody declined gradually with cloning. This problem was not improved in spite of our efforts with amounts of antibody and kinds of adjuvant as well as culturing conditions. Condensation reaction between baccacin III and an activated ester (viniyl ester) of C13-carboxylic acid was attempted under a crude antibody as a preliminary experiment. Further examination of the antibody reaction has not yet been conducted owing to a loss of antibody production function.