1. CAR-T Cells for Cancer Treatment: Current Design and Next Frontiers
Virgínia Picanço-Castro, Kamilla Swiech, Kelen Cristina Ribeiro Malmegrim de Farias, and Dimas Tadeu Covas
Part I: Design of Chimeric Antigen Receptors and Vector Production
2. Selection of Antibody Fragments for CAR-T Cell Therapy from Phage Display Libraries
Nestor F. Leyton-Vastro, Marcelo M. Brigido, and Andrea Q. Maranhão
3. Phase I/II Manufacture of Lentiviral Vectors under GMP in an Academic Setting
Anindya Dasgupta, Stuart Tinch, Kathleen Szczur, Rebecca Ernst, Nathaniel Shryock, Courtney Kaylor, Kendall Lewis, Eric Day, Timmy Truong, and William Swaney
4. Production of Lentivirus for the Establishment of CAR-T Cells
Marlous G. Lana and Bryan E. Strauss
5. Optimized Production of Lentiviral Vectors for CAR-T Cell
Pablo Diego Moço, Mário Soares de Abreu Neto, Daianne Maciely Carvalho Fantacini, andVirgínia Picanço-Castro
6. Lentiviral Vectors Production in Suspension Culture Using Serum-Free Medium for the Transduction of CAR-T Cells
Aline Do Minh, Michelle Yen Tran, and Amine A. Kamen
Part II: CAR-T Cells Generation and Manufacturing
7. In vitro Transcribed (IVT)-mRNA CAR Therapy Development
Miliotou N. Androulla and Papadopoulou C. Lefkothea
8. Generation of Chimeric Antigen Receptor T Cells Using Gammaretroviral Vectors
Feiyan Moand Maksim Mamonkin
9. Generation of CAR+ T Lymphocytes using the Sleeping Beauty Transposon System
Leonardo Chicaybam, Luiza Abdo, and Martín H. Bonamino
10. Platforms for Clinical-grade Car-T Cell Expansion
Amanda Mizukamiand Kamilla Swiech
11. CAR-T Cell Expansion in a Xuri Cell Expansion System W25
Trevor A. Smith
12. Methods and Process Optimization for Large Scale CAR T Expansion Using the G-Rex Cell Culture Platform
Josh Ludwig and Mark Hirschel
Part III: CAR-T Cells Characterization and Quality Control
13. Replication Competent Lentivirus Analysis of Vector Transduced T Cell Products used in Cancer Immunotherapy Clinical Trials
Kenneth Cornetta, Sue Koop, Emily Nance, Kimberley House, and Lisa Duffy
14. Immunophenotypic Analysis of CAR-T Cells
Júlia Teixeira Cottas de Azevedo, Amanda Mizukami, Pablo Diego Moço and Kelen Cristina Ribeiro Malmegrim
15. Approaches of T Cell Activation and Differentiation for CAR-T Cell Therapies
Robert D. Schwab, Darel Martínez Bedoya,Tiffany R. King, Bruce L. Levine, and Avery D. Posey, Jr.
16. Determination of Cytotoxic Potential of CAR-T Cells in Co-cultivation Assays
Renata Silvestre Nacasaki, Pablo Diego Moço, and Virgínia Picanço-Castro
17. Analysis of CAR-mediated Tonic Signaling
Hugo Calderon, Maksim Mamonkin, and Sonia Guedan
18. Generation of Tumor Cells Expressing Firefly Luciferase (fLuc) to Evaluate the Effectiveness of CAR in a Murine Model
Marcelo de Souza Fernandes Pereira, Daianne Maciely Carvalho Fantacini, and Virginia Picanço e Castro
19. Analysis of Antitumor Effects of CAR-T Cells in Mice with Solid Tumors
Alba Rodriguez-Garcia, Keisuke Watanabe, and Sonia Guedan
This volume providescomprehensive 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.