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Inverse electron demand Diels–Alder click chemistry for pretargeted PET imaging and radioimmunotherapy
Nature Protocols ( IF 14.8 ) Pub Date : 2021-06-14 , DOI: 10.1038/s41596-021-00540-2
Samantha M Sarrett 1, 2 , Outi Keinänen 1, 3, 4 , Eric J Dayts 1 , Guillaume Dewaele-Le Roi 1, 3, 5 , Cindy Rodriguez 1, 5 , Kathryn E Carnazza 6 , Brian M Zeglis 1, 2, 3, 5, 7
Affiliation  

Radiolabeled antibodies have shown promise as tools for both the nuclear imaging and endoradiotherapy of cancer, but the protracted circulation time of radioimmunoconjugates can lead to high radiation doses to healthy tissues. To circumvent this issue, we have developed an approach to positron emission tomography (PET) imaging and radioimmunotherapy (RIT) predicated on radiolabeling the antibody after it has reached its target within the body. This in vivo pretargeting strategy is based on the rapid and bio-orthogonal inverse electron demand Diels–Alder reaction between tetrazine (Tz) and trans-cyclooctene (TCO). Pretargeted PET imaging and RIT using TCO-modified antibodies in conjunction with Tz-bearing radioligands produce high activity concentrations in target tissues as well as reduced radiation doses to healthy organs compared to directly labeled radioimmunoconjugates. Herein, we describe how to prepare a TCO-modified antibody (humanized A33-TCO) as well as how to synthesize two Tz-bearing radioligands: one labeled with the positron-emitting radiometal copper-64 ([64Cu]Cu-SarAr-Tz) and one labeled with the β-emitting radiolanthanide lutetium-177 ([177Lu]Lu-DOTA-PEG7-Tz). We also provide a detailed description of pretargeted PET and pretargeted RIT experiments in a murine model of human colorectal carcinoma. Proper training in both radiation safety and the handling of laboratory mice is required for the successful execution of this protocol.



中文翻译:

用于预靶向 PET 成像和放射免疫疗法的逆电子需求 Diels-Alder 点击化学

放射性标记抗体已显示出作为癌症核成像和放射内放射治疗工具的前景,但放射免疫偶联物的循环时间延长会导致健康组织受到高辐射剂量。为了规避这个问题,我们开发了一种正电子发射断层扫描 (PET) 成像和放射免疫疗法 (RIT) 方法,该方法基于在抗体到达体内目标后对其进行放射性标记。这种体内预靶向策略基于四嗪 (Tz) 和反式之间的快速和生物正交逆电子需求 Diels-Alder 反应-环辛烯 (TCO)。与直接标记的放射免疫偶联物相比,使用 TCO 修饰的抗体和带有 Tz 的放射性配体的预靶向 PET 成像和 RIT 在靶组织中产生高活性浓度,并减少对健康器官的辐射剂量。在这里,我们描述了如何制备 TCO 修饰抗体(人源化 A33-TCO)以及如何合成两种带有 Tz 的放射性配体:一种用正电子发射放射性金属铜 64([ 64 Cu]Cu-SarAr- Tz) 和一种用发射 β 的放射性镧系元​​素 lutetium-177 ([ 177 Lu]Lu-DOTA-PEG 7-Tz)。我们还详细描述了人类结肠直肠癌小鼠模型中的预靶向 PET 和预靶向 RIT 实验。成功执行该协议需要在辐射安全和实验室小鼠处理方面进行适当的培训。

更新日期:2021-06-14
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