Comparative analysis and optimization of protocols for producing recombinant lentivirus carrying the anti-Her2 chimeric antigen receptor gene

The Journal of Gene Medicine
Weihua YuanHongzhi Li

Abstract

The production of anti-Her2 chimeric antigen receptor (CAR) T cells needs to be optimized to make it a reliable therapy. Three types of lentiviral vectors expressing anti-Her2 CAR together with packaging plasmids were co-transfected into 293 T-17 cells. The vector with the best packaging efficiency was selected, and the packaging cell culture system and packaging plasmid system were optimized. Centrifugation speed was optimized for the concentration of lentivirus stock. The various purification methods used included membrane filtration, centrifugation with a sucrose cushion and the novelly-designed instantaneous high-speed centrifugation. The recombinant lentiviruses were transduced into human peripheral T cells with an optimized multiplicity of infection (MOI). CAR expression levels by three vectors and the efficacy of CAR-T cells were compared. When co-transfected, packaging cells in suspension were better than the commonly used adherent culture condition, with the packaging system psPAX2/pMD2.G being better than pCMV-dR8.91/pVSV-G. The optimal centrifugation speed for concentration was 20 000 g, rather than the generally used ultra-speed. Importantly, adding instantaneous centrifugation for purification significantly increas...Continue Reading

References

Dec 19, 2002·Physiological Genomics·Jason E ColemanMohan K Raizada
Nov 16, 2004·Journal of Virological Methods·Miguel Sena-EstevesAlan W Flake
Nov 18, 2005·The Journal of Pharmacology and Experimental Therapeutics·Tzu-Hsuan Huang, Sherie L Morrison
Feb 2, 2006·Biotechnology Advances·María de Las Mercedes SeguraAlain Garnier
Mar 23, 2006·Clinical Cancer Research : an Official Journal of the American Association for Cancer Research·Pia WülfingBurkhard Brandt
Sep 2, 2006·Proceedings of the National Academy of Sciences of the United States of America·Kum-Joo ShinIain D C Fraser
Oct 26, 2006·Clinical Cancer Research : an Official Journal of the American Association for Cancer Research·Michael H KershawPatrick Hwu
Jan 19, 2007·Annals of Oncology : Official Journal of the European Society for Medical Oncology·G ValabregaM Aglietta
Apr 29, 2008·Annals of Oncology : Official Journal of the European Society for Medical Oncology·C Gravalos, A Jimeno
Jul 2, 2008·Neoplasia : an International Journal for Oncology Research·Stefan K SiwkoYi Li
Dec 31, 2008·Journal of Virological Methods·Hitoshi KurodaJakob Reiser
Mar 21, 2009·Nature Protocols·Robert H KutnerJakob Reiser
Sep 21, 2010·Seminars in Cell & Developmental Biology·Zheng CaiMark I Greene
Nov 9, 2010·Current Gene Therapy·M Schweizer, O-W Merten
Aug 31, 2011·Obstetrics and Gynecology International·Karim S Elsahwi, Alessandro D Santin
Jan 21, 2012·The Journal of Gene Medicine·Kevin J CurranRenier J Brentjens
Mar 31, 2012·Science Translational Medicine·Steven A Rosenberg
Apr 20, 2012·Immunotherapy·Christina D Pham, Duane A Mitchell
Feb 21, 2014·Science Translational Medicine·Marco L DavilaRenier Brentjens
Dec 2, 2014·Oncology Letters·Yong TangBarbara C Vanderhyden
Jan 5, 2017·Journal of Hematology & Oncology·Erhao Zhang, Hanmei Xu
Mar 24, 2017·Human Gene Therapy Methods·Olivia WilkinsTerence R Flotte
Sep 8, 2017·JAMA Oncology·John M Pagel, Howard Jack West
Nov 10, 2017·International Journal of Cancer. Journal International Du Cancer·Maria-Luisa SchubertMichael Schmitt

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