受现场自组装原理的启发，酶活化的分子纳米探针的开发对于靶向肿瘤检测具有深远影响。然而，尽管其固有优势，但在原始环境中获得高特异性的酶活性光学读出仍然具有挑战性。在这里，报告了一种全新的基于拉曼活性的自组装生物正交酶识别（nanoSABER）探针类别，用于靶向肿瘤成像。这类拉曼探针呈现狭窄的光谱带，反映其振动指纹，并为在不同的生物组织级别进行光学成像提供了一种有吸引力的解决方案。光学信标利用酶响应肽序列、独特的穿透肿瘤性质以及具有在细胞静默拉曼窗口的伸缩频率的振动标签。nanoSABER的设计经过了定制和工程化，可在目标酶存在的情况下转变为一种超分子结构，并展示出明显的振动特征，从而在酶活性和拉曼信号之间创建了直接的因果关系。通过与肿瘤相关酶胰蛋白酶专一底物结合，展示了nanoSABER在分子、细胞和组织水平上成像酶活性的独特能力，结合机器学习模型。这些结果表明，nanoSABER探针可能成为一种用于基于拉曼的识别肿瘤侵袭性、药物积累和治疗反应的多功能平台。©2023年作者。由Wiley-VCH GmbH出版的《Advanced Science》发表。

Inspired by the principle of in situ self-assembly, the development of enzyme-activated molecular nanoprobes can have a profound impact on targeted tumor detection. However, despite their intrinsic promise, obtaining an optical readout of enzyme activity with high specificity in native milieu has proven to be challenging. Here, a fundamentally new class of Raman-active self-assembling bioorthogonal enzyme recognition (nanoSABER) probes for targeted tumor imaging is reported. This class of Raman probes presents narrow spectral bands reflecting their vibrational fingerprints and offers an attractive solution for optical imaging at different bio-organization levels. The optical beacon harnesses an enzyme-responsive peptide sequence, unique tumor-penetrating properties, and vibrational tags with stretching frequencies in the cell-silent Raman window. The design of nanoSABER is tailored and engineered to transform into a supramolecular structure exhibiting distinct vibrational signatures in presence of target enzyme, creating a direct causality between enzyme activity and Raman signal. Through the integration of substrate-specific for tumor-associated enzyme legumain, unique capabilities of nanoSABER for imaging enzyme activity at molecular, cellular, and tissue levels in combination with machine learning models are shown. These results demonstrate that the nanoSABER probe may serve as a versatile platform for Raman-based recognition of tumor aggressiveness, drug accumulation, and therapeutic response.© 2023 The Authors. Advanced Science published by Wiley-VCH GmbH.