Real-Time Monitoring of CAR‑T Cell Efficiency through a Biorthogonal Cycloaddition Labeling Strategy

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 cyt...

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Bibliographic Details
Published in:Bioconjugate chemistry 2023-02, Vol.34 (2), p.443-452
Main Authors: Teng, Muzhou, Li, Zhijia, Zhou, Yali, Zhang, Zhengchao, Miao, Lele, Bai, Xiao, Li, Yumin, Wang, Song
Format: Article
Language:English
Subjects:
Alkynes
Antigen (tumor-associated)
Antigens
Antigens, Neoplasm
Cell membranes
Cell surface
Chimeric antigen receptors
Cycloaddition
Cycloaddition Reaction
Cytotoxicity
Fluorescence
Green fluorescent protein
Humans
Labels
Lymphocytes
Lymphocytes T
Major histocompatibility complex
Neoplasms
T-Lymphocytes
Tumor cells
Tumors
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Summary: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.
ISSN:1043-1802
1520-4812
DOI:10.1021/acs.bioconjchem.3c00006