Basic procedures for detection and cytotoxicity of chimeric antigen receptors

Keichiro Mihara, Tetsumi Yoshida, Joyeeta Bhattacharyya

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Citation (Scopus)

Abstract

Chimeric antigen receptors against CD19 (anti-CD19-CAR) are widely recognized and used by not only researchers associated with immunology, molecular biology, and cell biology but also physicians to treat B-cell malignancies. Anti-CD19-CAR is currently clinically available as one of the therapeutic modalities for refractory acute B-cell-typed lymphoblastic leukemia (B-ALL) patients. However, to detect CAR on the cell surface and investigate the efficacy of CAR-T cells, there are numerous experimental modalities including flow cytometry, the Cr-releasing assay, immunoblot, and immunostaining. We have chosen several techniques, which are necessary and sufficient as well as reliable and reproducible to detect and assess the killing effect of CAR-T cells. Here, we describe protocols for basic experiments and procedures for the detection of CAR on transduced cells and in in vitro coculture experiments to assess cytotoxicity using CAR-T cells.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages299-306
Number of pages8
DOIs
Publication statusPublished - 01-01-2019
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume1904
ISSN (Print)1064-3745

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

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    Mihara, K., Yoshida, T., & Bhattacharyya, J. (2019). Basic procedures for detection and cytotoxicity of chimeric antigen receptors. In Methods in Molecular Biology (pp. 299-306). (Methods in Molecular Biology; Vol. 1904). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-8958-4_13