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目的探讨锝99m(99mTc)标记抗烟曲霉单抗Mab-WF-AF-1的方法,观察标记物体外特性以及在正常小鼠体内生物学分布的特点。方法采用间接标记方法进行标记,薄层层析法检测标记物的标记率和稳定性,体外实验对标记物免疫活性进行鉴定。正常小鼠尾静脉注射3.7 MBq标记物,注射后40 min、2 h、4 h、7 h后取各器官称重后测放射性计数,评价标记物在正常小鼠体内生物分布特性。结果99mTc-WF-AF-1标记率达到95%以上,在磷酸盐缓冲液和血清中稳定性较高。标记物与烟曲霉的结合显著高于金黄色葡萄球菌[(10.32±0.21)%比(1.88±0.45)%,P<0.05]和白假丝酵母菌[(10.32±0.21)%比(1.67±0.29)%,P<0.05]。加入过量未标记蛋白后,标记物与烟曲霉孢子的结合显著降低。生物分布实验表明,标记物主要分布在肝、脾和肾,40 min和7 h的血池放射活性分别为(2.51±0.23)%ID/g和(0.53±0.13)%ID/g。结论99mTc标记Mab-WF-AF-1制备方法简便,标记率和放化纯度较高,稳定性好,标记物在小鼠体内主要通过肝脏和肾脏代谢,血液清除较快。
Objective To investigate the method of technetium 99m (99mTc) labeled anti-A. fumigatus monoclonal antibody Mab-WF-AF-1 to observe the in vitro characteristics of the marker and its biological characteristics in normal mice. Methods Indirect labeling method was used for labeling. The labeling rate and stability of the marker were detected by TLC. The immunological activity of the marker was identified in vitro. Normal mice were injected with 3.7 MBq marker through caudal vein, and the radioactivity was measured after 40 min, 2 h, 4 h and 7 h after injection, and the biodistribution of the marker in normal mice was evaluated. Results The labeling rate of 99mTc-WF-AF-1 reached more than 95% and its stability in phosphate buffer and serum was high. The binding of the marker to Aspergillus fumigatus was significantly higher than that of S. aureus [(10.32 ± 0.21)% (1.88 ± 0.45)%, P <0.05] and Candida albicans 0.29)%, P <0.05]. After addition of excess unlabeled protein, the binding of the marker to Aspergillus fumigatus spores was significantly reduced. Biodistribution experiments showed that the markers were mainly distributed in the liver, spleen and kidney. The radioactivity in the blood pool at 40 min and 7 h was (2.51 ± 0.23)% ID / g and (0.53 ± 0.13)% ID / g, respectively. Conclusion The 99mTc-labeled Mab-WF-AF-1 is simple, has high labeling and radiochemical purity, and has good stability. The marker is mainly metabolized by the liver and kidney in mice and the blood is cleared rapidly.