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Updated: Jun 26, 2026
A Real-time Potency Assay for Chimeric Antigen Receptor T Cells Targeting Solid and Hematological Cancer Cells
Published on: November 12, 2019
Intein-based modular chimeric antigen receptor platform for specific CD19/CD20 co-targeting
Pablo Gonzalez-Garcia1,2, Noelia Moares1,2, Wenjie Yi-He1,2
1Departmento de Biomedicina, Biotecnología y Salud Publica, Facultad de Medicina, Universidad de Cadiz, Spain.
Abstract:
Development of chimeric antigen receptor T-cell therapy has revolutionized the treatment of B-cell malignancies, although challenges such as antigen escape and tumor heterogeneity often decrease treatment success. Modular CARs targeting multiple antigens have been proposed as an interesting solution to address these challenges by reducing the likelihood of tumor cells evading treatment through the loss of a single antigen. In this study, we present a new modular CAR platform, termed CARtein, which takes advantage of intein interactions to jointly target CD19 and CD20 antigens. We demonstrate that the CARtein system, which features a universal CAR signaling backbone that covalently binds to specific scFv-intein recognition partners, generates fully active CARs. Functionality was validated using Raji cells and K562 cells expressing CD19 and/or CD20, observing significant T cell activation through NFAT and NFκB promoter activity and CD69 upregulation. Overall, our study lays the foundation for the establishment of a new way to target multiple antigens through a universal and inert CAR backbone with highly specific activation.
Insights
CARtein, a novel modular chimeric antigen receptor (CAR) platform, effectively targets multiple antigens like CD19 and CD20. This innovative approach enhances T-cell activation, addressing challenges in B-cell malignancy treatment.
Area of Science:
- Immunology
- Oncology
- Biotechnology
Background:
- Chimeric antigen receptor (CAR) T-cell therapy has transformed B-cell malignancy treatment.
- Challenges like antigen escape and tumor heterogeneity limit CAR T-cell efficacy.
- Modular CARs offer a strategy to overcome these limitations by targeting multiple antigens.
Purpose of the Study:
- To introduce CARtein, a novel modular CAR platform utilizing intein interactions.
- To enable simultaneous targeting of CD19 and CD20 antigens with a universal CAR backbone.
- To establish a foundation for a versatile multi-antigen targeting system.
Main Methods:
- Development of the CARtein platform with a universal CAR signaling backbone and scFv-intein recognition partners.
- Validation of CARtein functionality using CD19 and/or CD20 expressing Raji and K562 cells.
- Assessment of T-cell activation via NFAT and NFκB promoter activity and CD69 expression.
Main Results:
- The CARtein system successfully generated fully active CARs.
- Significant T-cell activation was observed when targeting cells expressing CD19 and/or CD20.
- Demonstrated specific activation through covalent binding of scFv-intein partners to the CAR backbone.
Conclusions:
- CARtein provides a new modular CAR platform for multi-antigen targeting in B-cell malignancies.
- The system utilizes intein-mediated covalent binding for precise CAR activation.
- This approach holds promise for overcoming antigen escape and improving CAR T-cell therapy outcomes.
