阴离子脂质体介导的腺病毒/卡莫司汀共转运系统载药量测定

目的建立一套有效可靠的检测方法,以测定阴离子脂质体介导的腺病毒和化疗药卡莫司汀共转运载体系统中腺病毒和卡莫司汀的含量。方法以钙离子融合法制备共转运载体复合物;根据腺病毒(Ad5)的hexon基因序列设计用于Taqman探针荧光定量聚合酶链式反应(PCR)的引物和探针序列,优化反应体系,从而准确测定腺病毒含量;优化色谱条件,建立针对共转运载体复合物检测卡莫司汀含量的高效液相色谱法。结果以荧光定量聚合酶链式反应法测得共转运载体复合物中腺病毒载药量为(23.2±1.8)%;高效液相色谱法测得卡莫司汀在共转运载体复合物中的载药量为(55±2.8)%。结论本实验成功建立了分别用于测定共转运载体系统中腺病毒和卡莫司汀含量的荧光定量聚合酶链式反应和高效液相色谱法;这两种检测方法操作简便、准确可靠,具有一定的普遍适用性。

Quantitative Determination of Adenovirus and Carmustine in Anionic Liposomes-mediated Co-Delivery System

OBJECTIVE To establish effective and reliable methods for the determination of adenovirus and carmustine in anionic liposomes-mediated co-delivery system. METHODS The complexes of co-delivery system were prepared by calcium-induced phase changing method. For carrying out the quantitative-PCR (Q-PCR) detection， the sequences of primer and probe were designed accoding to the hexon gene sequences of Ad5， and then the reaction system of Q-PCR was optimized to detect the loading rate of Ad5. The HPLC condition was also optimized to determine the drug loading of carmustine in the co-delivery system. RESULTS In the co-delivery system， the loading rate of adenovirus detected by Q-PCR method was （23.2±1.8）% and that of carmustine was （55±2.8）%. CONCLUSION Both of Q-PCR and HPLC methods have been successfully used for the quantification of adenovirus and carmustine in anionic liposomes-mediated co-delivery system. These methods are simple， reliable and accurate， which can be used in other similar experiments.