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Related Concept Videos

Reporter Genes02:11

Reporter Genes

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Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
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Related Experiment Video

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High Yield Expression of Recombinant Human Proteins with the Transient Transfection of HEK293 Cells in Suspension
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Updated transient gene expression protocol in Expi293F cells using PEI.

Gustaf Hederoth1, Andrés de la Rosa1, Ana Godec1

  • 1Department of Pharmacy, Uppsala University, Uppsala, Sweden.

Frontiers in Bioengineering and Biotechnology
|December 19, 2025
PubMed
Summary
This summary is machine-generated.

This study optimizes transient gene expression (TGE) for antibody production. Key improvements include reduced costs, faster workflows, and increased capacity, making therapeutic protein production more efficient.

Keywords:
Expi293FPEIcelpurelab-scaleprotocol updatesalmon sperm DNAtransfectiontransient gene expression protocol

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Area of Science:

  • Biotechnology
  • Protein Engineering
  • Biopharmaceutical Manufacturing

Background:

  • Transient gene expression (TGE) is vital for rapid protein therapeutic production, especially antibodies needing post-translational modifications.
  • Previous protocols enabled efficient TGE of complex antibodies.
  • Need for further optimization in cost, speed, and capacity for TGE workflows.

Purpose of the Study:

  • To present an optimized TGE protocol for enhanced antibody production.
  • To reduce costs associated with TGE media and plasmid DNA.
  • To accelerate workflow speed and increase production capacity.

Main Methods:

  • Replaced Expi293 medium with BalanCD HEK293 medium, reducing costs by ~90%.
  • Incorporated salmon sperm DNA as filler to minimize plasmid DNA usage.
  • Utilized diatomaceous earth (Celpure®) for rapid cell/debris sequestration, shortening harvest time to 15 minutes.
  • Recommended HiScreen Fibro PrismA columns for accelerated downstream purification.

Main Results:

  • Substantial cost reduction in media and plasmid DNA.
  • Harvesting time reduced from 2.5 hours to 15 minutes.
  • Streamlined workflow with maintained antibody yield and efficiency.
  • Enhanced scalability and throughput for TGE in Expi293F cells.

Conclusions:

  • The optimized TGE protocol significantly improves cost-effectiveness, speed, and capacity for antibody production.
  • These enhancements make TGE a more viable and efficient method for biopharmaceutical manufacturing.
  • The protocol facilitates scalable and high-throughput production of therapeutic proteins.