Leu J G, Chen B X, Schiff P B, Erlanger B F
Department of Pharmacology, College of Physicians and Surgeons, Columbia University, New York, New York 10032.
Cancer Res. 1993 Mar 15;53(6):1388-91.
Anti-taxol antibodies were generated in the rabbit using a taxol-bovine serum albumin conjugate prepared from 2'-succinyltaxol using a mixed anhydride procedure. Immunization with 2'-succinyltaxol-bovine serum albumin gave rise to polyclonal anti-taxol antibodies. By a radioimmunoassay using [3H]taxol, a standard curve gave a 50% inhibitory concentration of 1.0 nM. Taxol levels in human serum could be measured, with the lower limit of detection and measurement being 0.1 nM or 0.085 ng/ml. Two mouse monoclonal anti-taxol antibodies were isolated by immunizing BALB/c mice with the same antigen. One was an immunoglobulin G1 (69E4A8E) and the other was immunoglobulin M (29B7B3C). The specificity of these antibodies was determined by a competitive enzyme-linked immunosorbent assay with taxol and 10 different related derivatives and analogues. 29B7B3C had higher binding affinities for biologically active derivatives and markedly lower affinities for inactive derivatives; i.e., the specificity was consistent with the results of tubulin disassembly and cytotoxicity studies using the same taxol derivatives, making it suitable for screening for taxol or taxol-like compounds in extracts of natural products. 69E4A8E recognized the benzamidocarbamyl group at the C-3' position of taxol and had a lower affinity for other active compounds with different substitutions. Taxol levels in human serum could be detected and measured by 69E4A8E using a competitive enzyme-linked immunosorbent assay. The lower limit of measurement was about 50 nM or approximately 42 ng/ml. Similar measurements could be made by radioimmunoassay.
使用混合酸酐法由2'-琥珀酰紫杉醇制备的紫杉醇-牛血清白蛋白偶联物在兔体内产生抗紫杉醇抗体。用2'-琥珀酰紫杉醇-牛血清白蛋白免疫可产生多克隆抗紫杉醇抗体。通过使用[3H]紫杉醇的放射免疫测定，标准曲线给出的50%抑制浓度为1.0 nM。人血清中的紫杉醇水平可以被检测，检测和测量的下限为0.1 nM或0.085 ng/ml。用相同抗原免疫BALB/c小鼠，分离出两种小鼠单克隆抗紫杉醇抗体。一种是免疫球蛋白G1（69E4A8E），另一种是免疫球蛋白M（29B7B3C）。这些抗体的特异性通过与紫杉醇以及10种不同相关衍生物和类似物的竞争性酶联免疫吸附测定来确定。29B7B3C对生物活性衍生物具有更高的结合亲和力，对无活性衍生物的亲和力明显更低；即，其特异性与使用相同紫杉醇衍生物的微管蛋白解聚和细胞毒性研究结果一致，使其适用于筛选天然产物提取物中的紫杉醇或紫杉醇样化合物。69E4A8E识别紫杉醇C-3'位的苯甲酰胺基甲酰基，对其他具有不同取代基的活性化合物亲和力较低。使用竞争性酶联免疫吸附测定，69E4A8E可以检测和测量人血清中的紫杉醇水平。测量下限约为50 nM或约42 ng/ml。通过放射免疫测定也可以进行类似的测量。
