Key Laboratory of the Digestive System Tumors of Gansu Province, The Second Clinical Medical College of Lanzhou University, Lanzhou University Second Hospital, Lanzhou 730000, China.
Dermatology Hospital, Southern Medical University, Guangzhou 510091, China.
Bioconjug Chem. 2023 Feb 15;34(2):443-452. doi: 10.1021/acs.bioconjchem.3c00006. Epub 2023 Feb 7.
Chimeric antigen receptors (CARs) recognizing tumor-associated antigens (TAAs) effectively target tumor cells without using the major histocompatibility complex (MHC). However, CARs have inaccurate dose determination in clinical practice, and the methods that can solve this problem often produce cytotoxic substances, such as green fluorescent protein (GFP) insertion. Therefore, in this study, we tried to anchor harmless fluorescent labels on CAR-T cell membranes using highly biologically compatible strain-promoted alkyne-azide cycloaddition (SPAAC) without any byproducts. Our conjugated fluorescent label was stable on the CAR-T cell surface for at least two weeks, with excellent light stability and metrology. Also, this method enabled the rapid quantification of the living CAR-T cells without affecting their activity. Thus, this method is a promising reliable strategy for accurately diagnosing and treating cancer.
嵌合抗原受体 (CAR) 通过识别肿瘤相关抗原 (TAA) 来靶向肿瘤细胞，而无需使用主要组织相容性复合体 (MHC)。然而，CAR 在临床实践中的剂量确定并不准确，而能够解决此问题的方法通常会产生细胞毒性物质，例如绿色荧光蛋白 (GFP) 的插入。因此，在这项研究中，我们尝试使用高度生物兼容的应变促进叠氮-炔环加成 (SPAAC) 将无害的荧光标记物锚定在 CAR-T 细胞膜上，而不会产生任何副产物。我们的缀合荧光标记物在 CAR-T 细胞表面至少稳定两周，具有出色的稳定性和计量学性能。此外，该方法能够快速定量活的 CAR-T 细胞，而不会影响其活性。因此，该方法是一种很有前途的可靠策略，可用于准确诊断和治疗癌症。
