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This volume provides comprehensive methods from expert scientists working in the Chimeric Antigen Receptor T Cell (CAR-T Cell) field. Chapters guide readers through the state-of-art of CAR-T cell technology, CAR design and vector production, CAR-T cell generation and manufacturing, CAR-T cell characterization, and quality control. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, Chimeric Antigen Receptor T Cells: Development and Production aims to be useful in the production of CAR-T cells, especially for therapeutic purposes.
List of contents
CAR-T Cells for Cancer Treatment: Current Design and Next Frontiers.- Selection of Antibody Fragments for CAR-T Cell Therapy from Phage Display Libraries.- Phase I/II Manufacture of Lentiviral Vectors under GMP in an Academic Setting.- Production of Lentivirus for the Establishment of CAR-T Cells.- Optimized Production of Lentiviral Vectors for CAR-T Cell.- Lentiviral Vectors Production in Suspension Culture Using Serum-Free Medium for the Transduction of CAR-T Cells.- In vitro Transcribed (IVT)-mRNA CAR Therapy Development.- Generation of Chimeric Antigen Receptor T Cells Using Gammaretroviral Vectors.- Generation of CAR+ T Lymphocytes using the Sleeping Beauty Transposon System.- Platforms for Clinical-grade Car-T Cell Expansion.- CAR-T Cell Expansion in a Xuri Cell Expansion System W25.- Methods and Process Optimization for Large Scale CAR T Expansion Using the G-Rex Cell Culture Platform.- Replication Competent LentivirusAnalysis of Vector Transduced T Cell Products used in Cancer Immunotherapy Clinical Trials.- Immunophenotypic Analysis of CAR-T Cells.- Approaches of T Cell Activation and Differentiation for CAR-T Cell Therapies.- Determination of Cytotoxic Potential of CAR-T Cells in Co-cultivation Assays.- Analysis of CAR-mediated Tonic Signaling.- Generation of Tumor Cells Expressing Firefly Luciferase (fLuc) to Evaluate the Effectiveness of CAR in a Murine Model.- Analysis of Antitumor Effects of CAR-T Cells in Mice with Solid Tumors.
