Nakano Yukitaka, Saita Tetsuya, Fujito Hiroshi
Department of Pharmacy, Saga University Hospital, Japan.
Yakugaku Zasshi. 2012;132(6):727-32. doi: 10.1248/yakushi.132.727.
This paper reports a specific and sensitive enzyme-linked immunosorbent assay (ELISA) for pharmacokinetic studies of vindesine (VDS). Anti-VDS antibody was obtained by immunizing rabbits with VDS conjugated with bovine serum albumin using N-[β-(4-diazophenyl) ethyl] maleimide as a heterobifunctional coupling agent. An enzyme marker was similarly prepared by coupling VDS with horseradish peroxidase using N-(4-diazophenyl) maleimide. The detection limit of VDS by ELISA was approximately 24 pg/mL with 50-mL samples. This assay was specific for VDS and showed very slight cross-reactivity with other vinca alkaloids, vincristine (0.18%) and vinblastine (0.11%). The values for the VDS concentrations detected using this assay were comparable with those detected using HPLC. There was a good correlation between the values determined by the two methods. Moreover, ELISA was about 50-fold more sensitive in detecting VDS at lower concentrations. The sensitivity and specificity of ELISA should provide a useful tool for pharmacokinetic studies of VDS.
本文报道了一种用于长春地辛（VDS）药代动力学研究的特异性灵敏酶联免疫吸附测定法（ELISA）。使用N-[β-(4-重氮苯基)乙基]马来酰亚胺作为异双功能偶联剂，将长春地辛与牛血清白蛋白偶联后免疫家兔，获得抗长春地辛抗体。同样，使用N-(4-重氮苯基)马来酰亚胺将长春地辛与辣根过氧化物酶偶联，制备酶标记物。用ELISA法检测50 μL样品时，长春地辛的检测限约为24 pg/mL。该测定法对长春地辛具有特异性，与其他长春花生物碱长春新碱（0.18%）和长春碱（0.11%）的交叉反应性非常低。用该测定法检测的长春地辛浓度值与用高效液相色谱法检测的值相当。两种方法测定的值之间具有良好的相关性。此外，ELISA在检测较低浓度的长春地辛时灵敏度约高50倍。ELISA的灵敏度和特异性应为长春地辛的药代动力学研究提供有用的工具。
