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Large Volume Flow Electroporation of mRNA: Clinical Scale Process

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Book cover Synthetic Messenger RNA and Cell Metabolism Modulation

Part of the book series: Methods in Molecular Biology ((MIMB,volume 969))

Abstract

Genetic modification for enhancing cellular function has been continuously pursued for fighting diseases. Messenger RNA (mRNA) transfection is found to be a promising solution in modifying hematopoietic and immune cells for therapeutic purpose. We have developed a flow electroporation-based system for large volume electroporation of cells with various molecules, including mRNA. This allows robust and scalable mRNA transfection of primary cells of different origin. Here we describe transfection of chimeric antigen receptor (CAR) mRNA into NK cells to modulate the ability of NK cells to target tumor cells. High levels of CAR expression in NK cells can be maintained for 3–7 days post transfection. CD19-specific CAR mRNA transfected NK cells demonstrate targeted lysis of CD19-expressing tumor cells OP-1, primary B-CLL tumor cells, and autologous CD19+ B cells in in vitro assays with enhanced potency: >80% lysis at effector–target ratio of 1:1. This allows current good manufacturing practices (cGMP) and regulatory compliant manufacture of CAR mRNA transfected NK cells for clinical delivery.

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Acknowledgments

The authors acknowledge Dr. Dario Campana for the in-depth discussion and guidance in developing CAR mRNA technology. Thanks are also to Dr. James Brady for assistance with critical review of the manuscript.

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Authors and Affiliations

  1. MaxCyte Inc., Gaithersburg, MD, USA

    Linhong Li, Cornell Allen & Madhusudan V. Peshwa

  2. Max Cyte, Inc., Gaithersburg, MD, USA

    Rama Shivakumar

Authors
  1. Linhong Li
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  2. Cornell Allen
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  3. Rama Shivakumar
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  4. Madhusudan V. Peshwa
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Corresponding author

Correspondence to Madhusudan V. Peshwa .

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Editors and Affiliations

  1. The Anlyan Center, Department of Genetics, Yale School of Medicine, Cedar Street 300, New Haven, 06520-8005, Connecticut, USA

    Peter M. Rabinovich

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Li, L., Allen, C., Shivakumar, R., Peshwa, M.V. (2013). Large Volume Flow Electroporation of mRNA: Clinical Scale Process. In: Rabinovich, P. (eds) Synthetic Messenger RNA and Cell Metabolism Modulation. Methods in Molecular Biology, vol 969. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-260-5_9

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  • DOI: https://doi.org/10.1007/978-1-62703-260-5_9

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-259-9

  • Online ISBN: 978-1-62703-260-5

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