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Tuning the ignition of CAR: optimizing the affinity of scFv to improve CAR-T therapy

Abstract

How single-chain variable fragments (scFvs) affect the functions of chimeric antigen receptors (CARs) has not been well studied. Here, the components of CAR with an emphasis on scFv were described, and then several methods to measure scFv affinity were discussed. Next, scFv optimization studies for CD19, CD38, HER2, GD2 or EGFR were overviewed, showing that tuning the affinity of scFv could alleviate the on-target/off-tumor toxicity. The affinities of scFvs for different antigens were also summarized to designate a relatively optimal working range for CAR design. Last, a synthetic biology approach utilizing a low-affinity synthetic Notch (synNotch) receptor to achieve ultrasensitivity of antigen-density discrimination and murine models to assay the on-target/off-tumor toxicity of CARs were highlighted. Thus, this review provides preliminary guidelines of choosing the right scFvs for CARs.

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Acknowledgements

The authors would like to acknowledge the published work of many researchers whose work has relevance for this review, but were not cited because of space and our own limitations.

Funding

This research was funded by the National Natural Science Foundation of China Grants 31971324 (J.S.), 81973993 (X.G.) and 31971125 (C.Z.), by Zhejiang Provincial Natural Science Foundation Grants LR20H160003 (J.S.) and LY21H080002 (C.L.) and by the Pediatric Leukemia Diagnostic and Therapeutic Technology Research Center of Zhejiang Province JBZX-201904 (C.L.).

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Y.D., R.C. and J.S. wrote the manuscript. All the authors discussed and approved the manuscript.

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Correspondence to Jie Sun.

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Duan, Y., Chen, R., Huang, Y. et al. Tuning the ignition of CAR: optimizing the affinity of scFv to improve CAR-T therapy. Cell. Mol. Life Sci. 79, 14 (2022). https://doi.org/10.1007/s00018-021-04089-x

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  • DOI: https://doi.org/10.1007/s00018-021-04089-x

Keywords

  • Chimeric antigen receptor
  • scFv
  • Affinity
  • On-target/off-tumor
  • Adoptive cell therapy
  • Immunotherapy