{
  "abstract": "Background Chimeric Antigen Receptor (CAR) T cell manufacturing is associated with high costs. LipidBrick® Cell Ready is a novel lipid-based nanoparticle solution that can be easily loaded with mRNA and DNA payloads for transfecting T cells, without the need for specialized instruments.Methods Peripheral blood derived T cells were seeded into stirred bioreactor and activated. MODDE ®’s Design of Experiments (DOE) methodology was used to optimize the generation of T cell receptor (TCR) knockout (KO) T cells using the LipidBrick® Cell Ready lipid particles in static transfections.Results Activation induced robust T cell proliferation and stirring had no impact on cell growth. DOE guided optimization of KO efficiencies and T cell expansion identified optimal setpoints. Using these setpoints, TCR KO efficiencies >80% (pre-purification) were reproducibly observed across multiple donors, while maintaining cell growth.Conclusions Through DOE-guided optimization, we efficiently manufactured TCR KO T cells using significantly lower RNA to what is typically required in electroporation based-processes, thus simplifying manufacture and mitigating costs.Ethics Approval This study was approved by Sartorius’s Institutional Biosafety Committee (IBC). Project No.: 2024-02. Project title: T cell process development.",
  "authors": [
    {
      "affiliations": [
        "Sartorius Stedim North America, Marlborough, MA, USA"
      ],
      "name": "Sai Arun Batra"
    },
    {
      "affiliations": [
        "Sartorius Stedim North America, Marlborough, MA, USA"
      ],
      "name": "Lucas Nik Reger"
    },
    {
      "affiliations": [
        "Sartorius Stedim North America, Bohemia, NY, USA"
      ],
      "name": "Benjamin Reed"
    },
    {
      "affiliations": [
        "Sartorius Stedim North America, Bohemia, NY, USA"
      ],
      "name": "Michael Roberto"
    },
    {
      "affiliations": [
        "Sartorius Polyplus SAS, Illkirch-Graffenstaden, Illkirch-Graffenstaden, France"
      ],
      "name": "Claire Gueguen"
    },
    {
      "affiliations": [
        "Sartorius Polyplus SAS, Illkirch-Graffenstaden, Illkirch-Graffenstaden, France"
      ],
      "name": "Mélodie Seiler"
    },
    {
      "affiliations": [
        "Sartorius Stedim North America, Marlborough, MA, USA"
      ],
      "name": "Garima Agrahari"
    },
    {
      "affiliations": [],
      "name": "Nathan Chaboche"
    },
    {
      "affiliations": [
        "Sartorius Stedim North America, Marlborough, MA, USA"
      ],
      "name": "Sunandan Saha"
    }
  ],
  "title": "201 Optimization of T-cell receptor knock out using a novel lipid-based nanoparticle solution for the manufacture of allogeneic chimeric antigen receptor T cells",
  "uid": "50926a87-bd08-57d3-a7c0-b6dff783eea6"
}