Alirezapour Behrouz, Jalilian Amir Reza, Rajabifar Saeed, Mirzaii Mohammad, Moradkhani Sedigheh, Pouladi Mehraban, Aslani Gholamreza
Radiopharmacy Research Group, Radiation Application Research School, Nuclear Science and Technology Research Institute, 11365-3486, Tehran, Iran.
J Cancer Res Ther. 2014 Jan-Mar;10(1):112-20. doi: 10.4103/0973-1482.131434.
Herceptin and its fragments have been radiolabeled and used in the imaging of human epidermal growth factor receptor 2 (HER2)/neu-positive tumors and development of diagnostic kits is of great importance in radiopharmacy.
In this study, ¹¹¹ In-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid-trastuzumab (¹¹¹ In-DOTA-trastuzumab) was successively prepared and evaluated for ultimate use in the HER2 antigen imaging in oncology.
The conjugate was prepared, labeled and evaluated using in vitro (radioimmunoassay [RIA], enzyme-linked immunosorbent assay (ELISA), stability, binding, internalization)/in vivo (bio-distribution, single-photon emission computed tomography [SPECT]) experiments.
¹¹¹ In-DOTA-trastuzumab was prepared followed by determination of radiochemical purity (RCP), integrity of protein, immunoreactivity of radiolabeled antibody with HER2/neu antigen (by SkBr3 cell line binding and RIA methods) were determined followed by stability tests, internalization studies and the tissue bio-distribution determination in wild-type rats as well as SPECT imaging in SkBr3-bearing mice.
All values were expressed as mean ± standard deviation (mean ± SD) and the data were compared using Student's t-test. Statistical significance was defined as P < 0.05.
¹¹¹ In-DOTA-trastuzumab was prepared (RCP >95 ± 0.5%, S.A. 5.3 μCi/μg) with the average number of chelators per antibody of 6:1 showing significant immune-reactivity retention using ELISA. In vitro stability was >90% in phosphate buffered saline and 80 ± 0.5% in serum over 48 h. Cell binding was significant (>0.79). In vitro internalization reached up to %12-13 in 10 h. Significant tumor uptake was observed.
In vitro and in vivo/SPECT imaging in SkBr3-bearing mice demonstrated that ¹¹¹ In-DOTA-trastuzumab is a potential compound for molecular imaging of SPECT for diagnosis and follow-up of HER2 expression in oncology.
赫赛汀及其片段已被放射性标记，并用于人表皮生长因子受体2（HER2）/neu阳性肿瘤的成像，诊断试剂盒的开发在放射性药物领域具有重要意义。
在本研究中，成功制备了¹¹¹铟-1,4,7,10-四氮杂环十二烷-1,4,7,10-四乙酸-曲妥珠单抗（¹¹¹In-DOTA-曲妥珠单抗），并对其在肿瘤学HER2抗原成像中的最终应用进行了评估。
使用体外（放射免疫分析[RIA]、酶联免疫吸附测定[ELISA]、稳定性、结合、内化）/体内（生物分布、单光子发射计算机断层扫描[SPECT]）实验制备、标记并评估该偶联物。
制备¹¹¹In-DOTA-曲妥珠单抗，随后测定放射化学纯度（RCP）、蛋白质完整性、放射性标记抗体与HER2/neu抗原的免疫反应性（通过SkBr3细胞系结合和RIA方法），接着进行稳定性测试、内化研究以及野生型大鼠的组织生物分布测定和荷SkBr3小鼠的SPECT成像。
所有值均表示为平均值±标准差（mean±SD），数据采用学生t检验进行比较。统计学显著性定义为P<0.05。
制备了¹¹¹In-DOTA-曲妥珠单抗（RCP>95±0.5%，比活度5.3μCi/μg），每个抗体的螯合剂平均数量为6:1，使用ELISA显示出显著的免疫反应性保留。在磷酸盐缓冲盐水中48小时内体外稳定性>90%，在血清中为80±0.5%。细胞结合显著（>0.79）。10小时内体外内化率达到12%-13%。观察到显著的肿瘤摄取。
在荷SkBr3小鼠中的体外和体内/SPECT成像表明，¹¹¹In-DOTA-曲妥珠单抗是一种用于SPECT分子成像以诊断和随访肿瘤学中HER2表达的潜在化合物。
