Natarajan Arutselvan, Arksey Natasha, Iagaru Andrei, Chin Frederick T, Gambhir Sanjiv Sam
Mol Imaging. 2015;14. doi: 10.2310/7290.2014.00055.
Manufacturing of 64Cu-1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid (DOTA)-rituximab injection under good manufacturing practices (GMP) was validated for imaging of patients with CD20+ B-cell non-Hodgkin lymphoma. Rituximab was purified by size exclusion high performance liquid chromatography (HPLC) and conjugated to DOTA-mono-(N-hydroxysuccinimidyl) ester. 64CuCl2, buffers, reagents, and other raw materials were obtained as high-grade quality. Following a semi-automated synthesis of 64Cu-DOTA-rituximab, a series of quality control tests was performed. The product was further tested in vivo using micro-positron emission tomography/computed tomography (PET/CT) to assess targeting ability towards human CD20 in transgenic mice. Three batches of 64Cu-DOTA-rituximab final product were prepared as per GMP specifications. The radiolabeling yield from these batches was 93.1 ± 5.8%; these provided final product with radiopharmaceutical yield, purity, and specific activity of 59.2 ± 5.1% (0.9 ± 0.1 GBq of 64Cu), > 95% (by HPLC and radio-thin layer chromatography), and 229.4 ± 43.3 GBq/µmol (or 1.5 ± 0.3 MBq/µg), respectively. The doses passed apyrogenicity and human serum stability specifications, were sterile up to 14 days, and retained > 60% immunoreactivity. In vivo micro-PET/CT mouse images at 24 hours postinjection showed that the tracer targeted the intended sites of human CD20 expression. Thus, we have validated the manufacturing of GMP grade 64Cu-DOTA-rituximab for injection in the clinical setting.
按照药品生产质量管理规范（GMP）生产的64Cu-1,4,7,10-四氮杂环十二烷-N,N',N'',N'''-四乙酸（DOTA）-利妥昔单抗注射液，已通过验证，可用于CD20+B细胞非霍奇金淋巴瘤患者的成像。利妥昔单抗通过尺寸排阻高效液相色谱（HPLC）纯化，并与DOTA-单-（N-羟基琥珀酰亚胺）酯偶联。64CuCl2、缓冲液、试剂和其他原材料均为高品质产品。在半自动合成64Cu-DOTA-利妥昔单抗后，进行了一系列质量控制测试。该产品在体内使用微型正电子发射断层扫描/计算机断层扫描（PET/CT）进一步测试，以评估其对转基因小鼠体内人CD20的靶向能力。按照GMP规范制备了三批64Cu-DOTA-利妥昔单抗最终产品。这些批次的放射性标记产率为93.1±5.8%；最终产品的放射性药物产率、纯度和比活分别为59.2±5.1%（64Cu为0.9±0.1GBq）、>95%（通过HPLC和放射性薄层色谱法）和229.4±43.3GBq/µmol（或1.5±0.3MBq/µg）。这些剂量通过了无热原性和人血清稳定性规范，在14天内无菌，并保持>60%的免疫反应性。注射后24小时的体内微型PET/CT小鼠图像显示，该示踪剂靶向人CD20表达的预期部位。因此，我们已经验证了在临床环境中注射用GMP级64Cu-DOTA-利妥昔单抗的生产。
