Biomedical Engineering and Imaging Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA 02215-5418, USA.
STAR Protoc. 2021 Apr 5;2(2):100434. doi: 10.1016/j.xpro.2021.100434. eCollection 2021 Jun 18.
Noninvasive immunoimaging holds great potential for studying and stratifying disease as well as therapeutic efficacy. Radiolabeled single-domain antibody fragments (i.e., nanobodies) are appealing probes for immune landscape profiling, as they display high stability, rapid targeting, and excellent specificity, while allowing extremely sensitive nuclear readouts. Here, we present a protocol for radiolabeling an anti-CD11b nanobody and studying its uptake in mice by a combination of positron emission tomography imaging, gamma counting, and autoradiography. Our protocol is applicable to nanobodies against other antigens. For complete details on the use and execution of this protocol, please see Priem et al. (2020), Senders et al. (2019), or Rashidian et al. (2017).
非侵入性免疫成像在研究和分层疾病以及治疗效果方面具有巨大的潜力。放射性标记的单域抗体片段（即纳米体）是免疫景观分析的诱人探针，因为它们具有高稳定性、快速靶向和优异的特异性，同时允许进行极其敏感的核读出。在这里，我们提出了一种放射性标记抗 CD11b 纳米体的方案，并通过正电子发射断层扫描成像、γ计数和放射自显影的组合研究其在小鼠中的摄取。我们的方案适用于针对其他抗原的纳米体。有关此方案的使用和执行的完整详细信息，请参阅 Priem 等人（2020 年）、Senders 等人（2019 年）或 Rashidian 等人（2017 年）。
