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. 2024 Feb 5;26(1):24.
doi: 10.1208/s12248-023-00880-9.

Recommendations for Method Development and Validation of qPCR and dPCR Assays in Support of Cell and Gene Therapy Drug Development

Amanda Hays  1 , Mark Wissel  2 , Kelly Colletti  3 , Russell Soon  4 , Mitra Azadeh  5 , Justin Smith  6 , Rajitha Doddareddy  7 , Melanie Chalfant  8 , Wendy Adamowicz  9 , Swarna Suba Ramaswamy  10 , Sanjay L Dholakiya  11 , Sebastian Guelman  12 , Bryan Gullick  13 , Jennifer Durham  14 , Keith Rennier  15 , Pruthvi Nagilla  16 , Anamica Muruganandham  13 , Manisha Diaz  2 , Cassandra Tierney  6 , Kaarthik John  17 , Jenny Valentine  18 , Timothy Lockman  9 , Hsing-Yin Liu  19 , Benjamin Moritz  20 , Jean Paul Ouedraogo  21 , Marie-Soleil Piche  21 , Muriel Smet  22 , Jacqueline Murphy  19 , Kaylyn Koenig  23 , Agnes Zybura  15 , Carrie Vyhlidal  24 , Jonathan Mercier  25 , Niketa Jani  14 , Mikael Kubista  26 , Donald Birch  23 , Karlin Morse  23 , Oskar Johansson  27
Affiliations

Recommendations for Method Development and Validation of qPCR and dPCR Assays in Support of Cell and Gene Therapy Drug Development

Amanda Hays et al. AAPS J. .

Abstract

The emerging use of qPCR and dPCR in regulated bioanalysis and absence of regulatory guidance on assay validations for these platforms has resulted in discussions on lack of harmonization on assay design and appropriate acceptance criteria for these assays. Both qPCR and dPCR are extensively used to answer bioanalytical questions for novel modalities such as cell and gene therapies. Following cross-industry conversations on the lack of information and guidelines for these assays, an American Association of Pharmaceutical Scientists working group was formed to address these gaps by bringing together 37 industry experts from 24 organizations to discuss best practices to gain a better understanding in the industry and facilitate filings to health authorities. Herein, this team provides considerations on assay design, development, and validation testing for PCR assays that are used in cell and gene therapies including (1) biodistribution; (2) transgene expression; (3) viral shedding; (4) and persistence or cellular kinetics of cell therapies.

Keywords: AAV; RT-qPCR; biodistribution; cell therapy; cellular kinetics; dPCR; gene therapy; qPCR; shedding; transgene expression.

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References

    1. FDA. What is Gene therapy? https://www.fda.gov/vaccines-blood-biologics/cellular-gene-therapy-produ... . 25 July 2018. Accessed 25 June 2023.
    1. Sayed N, Allawadhi P, Khurana A, Singh V, Navik U, Pasumarthi SK, et al. Gene therapy: comprehensive overview and therapeutic applications. Life Sci. 2022;294: 120375. https://doi.org/10.1016/j.lfs.2022.120375 . - DOI - PubMed
    1. Jahangiri S, Rahimnejad M, Boroujeni NN, Ahmadi Z, Fath PM, Ahmadi S, et al. Viral and non-viral gene therapy using 3D (bio)printing. J Gene Med. 2022;24(12): e3458. https://doi.org/10.1002/jgm.3458 . - DOI - PubMed
    1. American Society of Gene and Cell Therapy. Gene & Cell Therapy FAQs. 2021. https://asgct.org/education/more-resources/gene-and-cell-therapy-faqs . Accessed 27 June 2023.
    1. Lin H, Cheng J, Mu W, Zhou J, Zhu L. Advances in universal CAR-T cell therapy. Front Immunol. 2021;12: 744823. https://doi.org/10.3389/fimmu.2021.744823 . - DOI - PubMed - PMC

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