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Selection of Antibody Fragments for CAR-T Cell Therapy from Phage Display Libraries
Nestor F Leyton-Castro1, Marcelo M Brigido1,2, Andrea Q Maranhão3,4
1Molecular Pathology Graduation Programme, School of Medicine, University of Brasilia, Brasilia, Brazil.
Methods in Molecular Biology (Clifton, N.J.)
|November 11, 2019
Summary
Researchers developed new methods to find novel single-chain variable fragments (scFv) for chimeric antigen receptors (CAR). These scFvs are crucial for CAR-T cell therapy advancements in cancer treatment.
Area of Science:
- Immunology
- Biotechnology
- Oncology
Background:
- Chimeric antigen receptor (CAR)-T cell therapy shows significant promise for cancer treatment.
- Developing novel CARs is essential to broaden the application of CAR-T cell therapy.
- The antigen-binding component of CARs is a single-chain variable fragment (scFv) that targets cancer-specific surface proteins.
Purpose of the Study:
- To discuss the utility of human scFv phage display libraries for identifying new monoclonal antibodies (mAbs) against tumor surface antigens.
- To present protocols for isolating novel scFvs for CAR construction.
Main Methods:
- Utilizing extracellular domains of surface proteins in biotinylated format as selection antigens.
- Employing elution with unlabeled peptides and selection in solution.
- Analyzing scFv enrichment using next-generation sequencing (NGS).
Main Results:
- Protocols were established for the isolation of scFvs targeting specific surface antigens.
- The described methods facilitate the identification of novel scFv binders.
Conclusions:
- The presented protocols enable the isolation of new scFvs.
- These scFvs can be utilized in constructing novel CARs for enhanced cancer therapy applications.
