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A Method For Production of Recombinant mCD1d Protein in Insect Cells.
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Insect High Five™ cell line development using site-specific flipase recombination technology.

Mafalda M Dias1,2, João Vidigal1,2, Daniela P Sequeira1,2

  • 1IBET, Instituto de Biologia Experimental e Tecnológica, Oeiras 2780-901, Portugal.

G3 (Bethesda, Md.)
|May 13, 2021
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Summary
This summary is machine-generated.

This study developed a stable insect cell line for biologics manufacturing using Flp recombinase-mediated cassette exchange and FACS. This new platform enables flexible, high-cell-density production of single or multiple proteins, accelerating development.

Keywords:
FACSRMCEinfluenza VLPsinsect Hi5 cellsstable expression

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Area of Science:

  • Biotechnology
  • Cell Biology
  • Molecular Biology

Background:

  • Insect High Five™ (Hi5) cells are utilized for heterologous protein production via baculovirus expression vector systems (BEVS) or transient gene expression.
  • Stable expression systems are needed to overcome limitations of current methods for large-scale biologics manufacturing, especially for high-cell-density processes.

Purpose of the Study:

  • To create a stable, master clonal Hi5 cell line for flexible, large-scale production of biologics.
  • To develop a system for expressing single or multiple proteins of interest from a tagged genomic locus.

Main Methods:

  • Combined Flp recombinase-mediated cassette exchange (RMCE) with fluorescence-activated cell sorting (FACS).
  • A three-step protocol involving random integration, selection, and FACS was used to generate the stable Hi5 cell line.
  • Demonstrated locus exchangeability with intracellular (iCherry) and extracellular (influenza HA and M1) protein expression models.

Main Results:

  • Successfully generated a stable Hi5 master cell line using RMCE and FACS.
  • The master clone demonstrated efficient and flexible expression of single and multiple proteins.
  • The developed platform facilitates high-cell-density culture for biologics manufacturing.

Conclusions:

  • The stable insect Hi5 cell platform offers a robust solution for biologics development.
  • This system enhances flexibility and efficiency in protein production for biopharmaceutical applications.
  • The platform has the potential to accelerate the development and manufacturing of biologics.