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piggyBac Transposon System Modification of Primary Human T Cells
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Recombinant CHO Cell Pool Generation Using PiggyBac Transposon System.

Sowmya Balasubramanian1,2

  • 1Laboratory of Cellular Biotechnology (LBTC), École Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland. balasubramanian.sowmya@gene.com.

Methods in Molecular Biology (Clifton, N.J.)
|June 26, 2024
PubMed
Summary

We developed a piggyBac transposon system to generate Chinese hamster ovary (CHO) cell pools for rapid recombinant protein production. This method ensures high titers and consistent product quality for biopharmaceutical manufacturing.

Keywords:
CHO cellsOrbital shakingRecombinant proteinTransfectionTransposon system

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Area of Science:

  • Biotechnology
  • Cell line development
  • Recombinant protein production

Background:

  • Chinese hamster ovary (CHO) cells are widely used for recombinant protein production.
  • Achieving high titers and consistent product quality is crucial for efficient biomanufacturing.
  • Rapid generation of stable cell pools is essential for timely protein production.

Purpose of the Study:

  • To describe a method for generating CHO cell pools with desirable characteristics for recombinant protein production.
  • To utilize the piggyBac transposon system for efficient gene integration in CHO cells.
  • To establish a robust platform for rapid biopharmaceutical manufacturing.

Main Methods:

  • Co-transfection of CHO cells with a donor plasmid (gene of interest) and a helper plasmid (transposase) using polyethyleneimine (PEI).
  • Employing the piggyBac transposon system for stable gene integration into the host cell genome.
  • Implementing a genetic selection strategy to isolate and expand desired cell pools.

Main Results:

  • Successful generation of CHO cell pools using the piggyBac transposon system.
  • Demonstrated high titer and consistent product quality in the generated cell pools.
  • Established a scalable method for rapid production of recombinant proteins.

Conclusions:

  • The piggyBac transposon system provides an efficient method for generating CHO cell pools for recombinant protein production.
  • The generated cell pools are suitable for initiating batch or fed-batch cultures.
  • This approach facilitates the rapid generation of clonal cell lines and cell banks for future biopharmaceutical manufacturing.