{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,15]],"date-time":"2026-09-15T03:02:39Z","timestamp":1789441359266,"version":"build-2803163510"},"reference-count":22,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2026,3,24]],"date-time":"2026-03-24T00:00:00Z","timestamp":1774310400000},"content-version":"vor","delay-in-days":23,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,3,1]],"date-time":"2026-03-01T00:00:00Z","timestamp":1772323200000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Current Protocols"],"published-print":{"date-parts":[[2026,3]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Chimeric antigen receptor (CAR)\u2010T cell treatment has shifted the paradigm of cancer immunotherapy, especially in B\u2010cell malignancies, with around an 80% response rate in acute lymphoblastic leukemia. Emerging data on the use of CAR\u2010T cells in the treatment of autoimmune diseases, such as lupus, indicate another significant advance in this therapeutic field and highlight its promising potential for the future. Despite its remarkable therapeutic potential, the manufacturing of CAR\u2010T cells, including the production of critical components such as high\u2010titer lentiviral vector (LV), remains a significant challenge, limiting the broader accessibility and the ease of manufacturing and administration\u00a0of this therapy. Lack of optimized and harmonized protocols makes the manufacturing of CAR\u2010T cells cumbersome. To ease some of the challenges of CAR\u2010T cell manufacturing, we optimized LV production, with an emphasis on enhancing viral titers using different transfer\u2010to\u2010helper plasmid ratios, optimizing transfection efficiency in HEK293T cells, and fine\u2010tuning peripheral blood mononuclear cell activation and transduction conditions. We present a set of optimized protocols for the scalable production of third\u2010generation LVs for CAR\u2010T cell generation. This approach facilitates preclinical and translational research by providing a framework for the reproducible, economical, and effective production of CAR\u2010T cells. \u00a9 2026 Wiley Periodicals LLC.<\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 1<\/jats:bold>\n                    : Plasmid ratio screening\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 2<\/jats:bold>\n                    : Viral concentration via ultracentrifugation\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 3<\/jats:bold>\n                    : FACS\u2010based titration assay\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 4<\/jats:bold>\n                    : Peripheral blood mononuclear cell (PBMC) isolation\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 5<\/jats:bold>\n                    : PBMC activation for effective transduction\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 6<\/jats:bold>\n                    : CAR\u2010T cell generation with lentivirus\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Support Protocol<\/jats:bold>\n                    : Flow cytometry gating strategies\n                  <\/jats:p>","DOI":"10.1002\/cpz1.70346","type":"journal-article","created":{"date-parts":[[2026,3,24]],"date-time":"2026-03-24T14:25:32Z","timestamp":1774362332000},"source":"Crossref","is-referenced-by-count":0,"title":["Optimized High\u2010Titer Lentivirus Production and Efficient CAR\u2010T Cell Generation"],"prefix":"10.1002","volume":"6","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-1174-2732","authenticated-orcid":false,"given":"Atul Kumar","family":"Roy","sequence":"first","affiliation":[{"name":"Centre for Immuno\u2010biology and Immunotherapy Translational Health Science and Technology Institute, Faridabad  Gurugram India"},{"name":"Centre for CAR\u2010T Cell Therapy Translational Health Science and Technology Institute, Faridabad  Gurugram India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ashwani","family":"Singh","sequence":"additional","affiliation":[{"name":"Centre for Immuno\u2010biology and Immunotherapy Translational Health Science and Technology Institute, Faridabad  Gurugram India"},{"name":"Centre for CAR\u2010T Cell Therapy Translational Health Science and Technology Institute, Faridabad  Gurugram India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jyotsna","family":"Dandotiya","sequence":"additional","affiliation":[{"name":"Centre for Immuno\u2010biology and Immunotherapy Translational Health Science and Technology Institute, Faridabad  Gurugram 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