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形成通常由紫外线或荧光胺方法证实。然而，由于2′-琥珀基紫杉醇与蛋白质的紫外吸收最大值相似，因此用这些方法很难证实2′-琥珀基紫杉醇与蛋白质的结合。因此，我们使用中性损失扫描和质谱/质谱技术进行了质谱分析，以确认2'-琥珀基紫杉醇蛋白偶联物的配方。以2′-琥珀基紫杉醇- rsa免疫Balb/c小鼠，每隔2周免疫一次。用常规方法处理小鼠脾细胞，得到4个产生抗紫杉醇单克隆抗体的杂交瘤。然而，杂交瘤产生抗紫杉醇单克隆抗体的能力不高，抗体滴度较低。不幸的是，随着克隆，抗体的产量逐渐下降。尽管我们对抗体的数量和佐剂的种类以及培养条件进行了努力，但这一问题并没有得到改善。在粗抗体条件下，尝试了baccacin III与c13 -羧酸活化酯（乙烯基酯）的缩合反应作为初步实验。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.