近年来，以肿瘤为诊断目的的近红外II荧光探针受到了广泛关注。在本研究中，使用 IRDye800CW-NHS 荧光染料与曲妥珠单抗构建了 NIR-II 荧光探针，并研究了其在异种移植小鼠模型中靶向 HER-2 阳性乳腺癌的能力。将该探针与使用类似结构合成的曲妥珠单抗-ICG（ICG-NHS）进行比较。结果表明，IRDye800CW-NHS 在 NIR-I 和 NIR-II 中的荧光明显强于 ICG-NHS 在水相中的荧光。并在生物成像实验中揭示了IRDye800CW-NHS和ICG-NHS不同的代谢模式。 IRDye800CW-NHS主要通过肾脏代谢，而ICG-NHS主要通过肝脏代谢。与曲妥珠单抗偶联后，基于计算的最大肿瘤肌肉比（TMR），曲妥珠单抗-800CW（TMR=5.35±0.39）不仅具有比曲妥珠单抗-ICG（TMR=4.42±0.10）更强的肿瘤靶向能力，而且还具有比曲妥珠单抗-ICG（TMR=4.42±0.10）更强的肿瘤靶向能力。肝脏摄取相对较低，新陈代谢较快。组织病理学分析证明两种荧光探针对多种器官组织均无毒。这些结果揭示了 IRDye800CW-NHS 优异的光学特性，以及与抗体偶联开发荧光探针的巨大潜力，有望应用于人类术中乳腺癌导航。

Near-infrared II fluorescent probes targeting tumors for diagnostic purposes have received much attention in recent years. In this study, a fluorescent probe for the NIR-II was constructed by using IRDye800CW-NHS fluorescent dye with Trastuzumab, which was investigated for its ability to target HER-2-positive breast cancer in xenograft mice models. This probe was compared with Trastuzumab-ICG which was synthesized using a similar structure, ICG-NHS. The results demonstrated that the IRDye800CW-NHS had significantly stronger fluorescence in the NIR-I and NIR-II than ICG-NHS in the aqueous phase. And the different metabolic modes of IRDye800CW-NHS and ICG-NHS were revealed in bioimaging experiments. IRDye800CW-NHS was mainly metabolised by the kidneys, while ICG-NHS was mainly metabolised by the liver. After coupling with Trastuzumab, Trastuzumab-800CW (TMR = 5.35 ± 0.39) not only had a stronger tumor targeting ability than Trastuzumab-ICG (TMR = 4.42 ± 0.10) based on the calculated maximum tumor muscle ratio (TMR), but also had a comparatively lower hepatic uptake and faster metabolism. Histopathology analysis proved that both fluorescent probes were non-toxic to various organ tissues. These results reveal the excellent optical properties of IRDye800CW-NHS, and the great potential of coupling with antibodies to develop fluorescent probes that will hopefully be applied to intraoperative breast cancer navigation in humans.