Expi293 Expression System

Trust the industry standard for rapid, high-yield transient HEK 293 expression

Every minute counts when it comes to expressing drug candidates. That’s why the Gibco Expi293 Expression System is the tried-and-true choice for transient expression in 293 cells. This rapid, high-yield system allows you to access recombinant 293–derived proteins in just five to seven days, so you get what you need faster.
 

Benefits of the Expi293 Expression System

Get more protein while reducing hands-on time and waste

Use your preferred culture format without sacrificing yield

Fully documented cGMP-banked cells

The graph represents the equivalent rabbit IgG titers are obtained in cGMP-banked Expi293F cells and catalog Expi293F cells at 30 mL scale using the Expi293 Expression System.

  • Fully documented cGMP-banked cell line 
  • Derived from the same Master Seed Bank as standard Expi293F Cells (Cat No. A14527
  • Pre-adapted to Expi293 Expression Medium 
  • A component of the fully integrated, animal origin-free Expi293 Expression System for applications requiring cGMP-banked cells 
  • Flexible commercial licensing options with no royalties

See product information

Get more protein while reducing hands-on time and waste

Use your preferred culture format without sacrificing yield

Fully documented cGMP-banked cells

The graph represents the equivalent rabbit IgG titers are obtained in cGMP-banked Expi293F cells and catalog Expi293F cells at 30 mL scale using the Expi293 Expression System.

  • Fully documented cGMP-banked cell line 
  • Derived from the same Master Seed Bank as standard Expi293F Cells (Cat No. A14527
  • Pre-adapted to Expi293 Expression Medium 
  • A component of the fully integrated, animal origin-free Expi293 Expression System for applications requiring cGMP-banked cells 
  • Flexible commercial licensing options with no royalties

See product information

What’s in the system?

The Expi293 Expression System brings together a high-expressing 293 cell line, a chemically defined serum–free culture medium, and a high-efficiency transfection reagent with specialized enhancers.

  • 2 vials of frozen Gibco Expi293F Cells
  • 1 L of Gibco Expi293 Expression Medium
  • 1 Gibco ExpiFectamine 293 Transfection Kit, sufficient to transfect 1 L of culture
  • Gibco Opti-MEM I Reduced Serum Medium
  • An antibody-expressing positive control vector

Additional capabilities of Expi293 Transient Protein Expression System for structural biology and inducible expression

Expi293 Transient Protein Expression System

Expect more from the system that transformed mammalian transient expression. Now introducing three additional cell lines and a labeling kit that unlock new possibilities within the Expi293 Transient Protein Expression System. With features accommodating structural biology and inducible protein expression applications, you can expand your work with the same incredible yield and intricate structure you’ve experienced with our other Expi293 Expression Systems. Get the high yields, rapid time to protein (just 5–7 days), and reproducibility you’ve come to expect, with the added flexibility to suit a broader range of applications and proteins.  

The Expi293 system’s additional capabilities include:

  • Glycan modulation
  • Inducible expression
  • Methionine protein labeling

Expi293F GnTI- Cell Line

With the Expi293F GnTI- Cell Line you no longer need to compromise yield to gain simplicity. These cells were designed to facilitate analysis techniques that depend on the glycosylation state of the protein.

Key benefits:

  • >30x higher yield than next-best performing product
  • Simplified glycan patterns
Bar chart showing amount of human IgG expressed in Expi293F GnTI(-) cells compared to competitor cells

Expi293F GnTI- cells express proteins at levels comparable to parental Expi293 cells. By comparison, expression levels of ATCC GnTI- cells adapted into FreeStyle293 media with PEI transfection are >30-fold lower than Expi293F GnTI- cells in the Expi293 System.

Western blot showing forms of CB2 GPCR protein expressed in Expi293F and in GnTI(-) cells

Expression of a GPCR in Expi293 (Lanes 1-4) and Expi293 GnTI- (Lanes 5-8) cells under varying expression conditions.

Expi293F Inducible GnTI- cell line

Expi293F Inducible GnTI- cell line you can achieve tunable expression of glycan modified proteins. These cells were designed to facilitate the analysis of complex proteins without adding workflow steps or sacrificing yield in your structural biology work and other applications.

Key benefits:

  • >8x higher yield over existing T-REx 293 cells
  • Tunable expression of proteins
  • Simplified glycan patterns
     
Bar chart showing amount of protein expressed in Expi293F GnTI(-) and TET-inducible cells compared to previous systems

Upon full induction in pcDNA5.0/TO vector, Expi293F Inducible GnTI- cells express proteins at levels comparable to parental Expi293F cells. By comparison, expression levels of T-REx 293 cells adapted into FreeStyle293 media with PEI transfection are 8-fold lower than the Expi293F Inducible GnTI- cell line in the Expi293 System.

Bar chart showing mean GFP fluorescence as a function of induction and multipanel brightfield and fluorescence images showing cell growth and fluorescence
imaging of tetracycline concentration

Expression of a GPCR with different levels of induction.

Glycan patterns for human IgG expressed in (A) Expi293F and (B) Expi293F GnTI- cell lines.

Expi293F Inducible cell line

The Expi293F Inducible cell line allows for regulated and tunable expression of proteins that may benefit from slower expression kinetics such as toxic proteins and membrane proteins. These cells were also designed to facilitate the study of cell models that require the regulation of protein expression.

Key benefits:

  • >9x higher yield than T-REx 293 cells
  • Tunable expression of proteins
Bar chart showing amount of protein expressed in Expi293F TET-inducible cells compared to previous systems

Upon full induction in pcDNA5.0/TO vector, Expi293F Tet-inducible cells express proteins at levels comparable to parental Expi293F cells. By comparison, expression levels of T-REx 293 cells adapted in FreeStyle293 media with PEI transfection are nearly 10-fold lower than the Expi293F Inducible cell line in the Expi293 System.

Bar chart showing mean titer as a function of induction via increasing tetracycline concentration

Cells were transfected with an Fc-fusion protein in pcDN5.0/TO vector and then induced with different levels of tetracycline in a dose-dependent manner.

Expi293F GnTI- Cell Line

With the Expi293F GnTI- Cell Line you no longer need to compromise yield to gain simplicity. These cells were designed to facilitate analysis techniques that depend on the glycosylation state of the protein.

Key benefits:

  • >30x higher yield than next-best performing product
  • Simplified glycan patterns
Bar chart showing amount of human IgG expressed in Expi293F GnTI(-) cells compared to competitor cells

Expi293F GnTI- cells express proteins at levels comparable to parental Expi293 cells. By comparison, expression levels of ATCC GnTI- cells adapted into FreeStyle293 media with PEI transfection are >30-fold lower than Expi293F GnTI- cells in the Expi293 System.

Western blot showing forms of CB2 GPCR protein expressed in Expi293F and in GnTI(-) cells

Expression of a GPCR in Expi293 (Lanes 1-4) and Expi293 GnTI- (Lanes 5-8) cells under varying expression conditions.

Expi293F Inducible GnTI- cell line

Expi293F Inducible GnTI- cell line you can achieve tunable expression of glycan modified proteins. These cells were designed to facilitate the analysis of complex proteins without adding workflow steps or sacrificing yield in your structural biology work and other applications.

Key benefits:

  • >8x higher yield over existing T-REx 293 cells
  • Tunable expression of proteins
  • Simplified glycan patterns
     
Bar chart showing amount of protein expressed in Expi293F GnTI(-) and TET-inducible cells compared to previous systems

Upon full induction in pcDNA5.0/TO vector, Expi293F Inducible GnTI- cells express proteins at levels comparable to parental Expi293F cells. By comparison, expression levels of T-REx 293 cells adapted into FreeStyle293 media with PEI transfection are 8-fold lower than the Expi293F Inducible GnTI- cell line in the Expi293 System.

Bar chart showing mean GFP fluorescence as a function of induction and multipanel brightfield and fluorescence images showing cell growth and fluorescence
imaging of tetracycline concentration

Expression of a GPCR with different levels of induction.

Glycan patterns for human IgG expressed in (A) Expi293F and (B) Expi293F GnTI- cell lines.

Expi293F Inducible cell line

The Expi293F Inducible cell line allows for regulated and tunable expression of proteins that may benefit from slower expression kinetics such as toxic proteins and membrane proteins. These cells were also designed to facilitate the study of cell models that require the regulation of protein expression.

Key benefits:

  • >9x higher yield than T-REx 293 cells
  • Tunable expression of proteins
Bar chart showing amount of protein expressed in Expi293F TET-inducible cells compared to previous systems

Upon full induction in pcDNA5.0/TO vector, Expi293F Tet-inducible cells express proteins at levels comparable to parental Expi293F cells. By comparison, expression levels of T-REx 293 cells adapted in FreeStyle293 media with PEI transfection are nearly 10-fold lower than the Expi293F Inducible cell line in the Expi293 System.

Bar chart showing mean titer as a function of induction via increasing tetracycline concentration

Cells were transfected with an Fc-fusion protein in pcDN5.0/TO vector and then induced with different levels of tetracycline in a dose-dependent manner.

Expi293 Met(–) Protein Labeling Kit

Use our Expi293 methionine-deficient protein labeling kit that streamlines structural biology studies by x-ray crystallography and NMR using high-titer mammalian suspension cultures.

Key benefits:

  • Uniform labeling of methionine residues
     
Bar charts of protein titer obtained from expression at different passages and growth conditions with methyl 13C methionine and selenomethionine and in the Expi293 expression system

(A) Protein expression with methyl 13C methionine added back at various time points prior to transfection. (B) Protein expression with selenomethionine added back at various time points prior to transfection.

Theoretical mass spectrographs showing peptides containing SeMet and a mix of native and SeMet-labeled peptide.

A comparison of theoretical spectra for peptide containing selenomethionine and the experimental spectra for a mixture of native and SeMet-labeled peptide.
 

Expi293 Expression System customer stories

Expi293 is cost-effective compared to other FreeStyle 293 Medium

NOTE: Cost comparison based on list price in USD and protein yields of 1 g/L with Expi293 system.

  PEI w/ FreeStyle 293 Medium Expi293 System (typically delivers 5x yield)
Cost of 1 L transfection $548 $1,531
Culture volume for equivalent yield 1 L 200 mL
Total cost for same amount of protein $607 $324

Ordering information

Thermo Scientific Nalgene sterile single-use Erlenmeyer flasks

Sterile, single-use Thermo Scientific Nalgene flasks are made from light, break-resistant, crystal-clear PETG, so you can express with confidence. All sizes are available in vented or non-vented formats, and with plain or baffled bottoms for use on the benchtop or shaker table.

See protocol for flasks validated with Expi293 Expression system