ExpiCHO Expression System | Thermo Fisher Scientific

Maintain protein quality and function from research to bioproduction

Switching from 293 to CHO cells during drug development doesn't have to create uncertainty. Now there's a better way. The Gibco ExpiCHO Expression System is a completely optimized system consisting of ExpiCHO-S cells that have been adapted to high-density, serum-free suspension culture in ExpiCHO Expression Medium, along with specially designed transfection reagents and enhancers, that provide high protein quality and quantity in a transient system. That means you can start your research work in CHO cells and stay in CHO cells through discovery.

“The ExpiCHO system helps speed up our developmental timelines by eliminating the need to have different hosts for transient versus stable expression. This eliminates a key quality risk when we transition from research into development.” Biotechnology company

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See the 5L flask designed for efficient ExpiCHO scale up

ExpiCHO protocol video and webcasts

Review these videos to learn how to produce recombinant proteins using the ExpiCHO Expression System and also to hear about the latest advances in protein expression from industry experts.

What’s in the ExpiCHO Expression System?

The Gibco ExpiCHO Expression System brings together a high-expressing CHO cell line, a chemically-defined animal origin-free, serum-free, and protein-free culture medium, an optimized culture feed, and a high-efficiency transfection reagent that work in concert to provide protein titers 3x higher than the Gibco Expi293 Expression System and 160x higher than the Gibco FreeStyle CHO Expression System. With the chemically-defined formulation of the ExpiCHO(TM) Expression Medium, you can come to expect high reproducibility in your results and lot-to-lot reliability.

2 vials of frozen Gibco ExpiCHO-S Cells
1 liter of Gibco ExpiCHO Expression Medium
1 Gibco ExpiFectamine CHO Transfection Kit, sufficient to transfect 1 liter of culture
Gibco OptiPRO Serum-Free Medium
An antibody-expressing positive control vector

Get up to 20x more protein in less volume

The ExpiCHO Expression System is designed to deliver superior protein yields compared to alternative transient systems, saving you precious time, incubator space, and plasticware costs. Now you can express the same amount of protein in a single flask that other transient CHO systems express in 20 or more flasks.

“The higher titers achieved by the ExpiCHO system… saves us considerable time, effort, and money." Bonnie Hammer, PhD, Vice President of Biologics Development, Invenra

Recombinant protein titers in ExpiCHO and other transient CHO system

Figure 1. Recombinant protein titers in ExpiCHO and other transient CHO expression systems. Expression levels of human IgG, rabbit IgG and erythropoietin in ExpiCHO and other transient CHO expression systems are shown. ExpiCHO titers range from 25–160 times those of other transient CHO expression systems.

Titer comparison of Expi293 and ExpiCHO Expression Systems

Expression levels of human IgG, rabbit IgG, murine IgG, and a broad range of non-Ab proteins in Expi293 or other transient expression systems and ExpiCHO systems are shown. IgG titers in ExpiCHO Expression System range from 0.6–97–fold (Avg. 5-fold) than those obtained using various other systems. Additionally, 7 proteins from this panel were not expressed in Expi293 cells yet expressed at high levels in the ExpiCHO system.

Human antibodies

Protein	Expi293 or current system (mg/L)	ExpiCHO (mg/L)	Fold increase
Human antibody	26.6	2,580	97
	4	147	33
	4	56	14.6
	22	266	12.4
	48	334	6.9
	36	190	5.3
	18	78	4.3
	85	314	3.7
	170	410	2.4
	138	325	2.4
	25	50	2.0
	102	204	2.0
	57	92	1.6
	336	491	1.5
	565	864	1.5
	450	600	1.3
	350	450	1.3
	309	383	1.2
	243	276	1.1
	352	371	1.1
	565	620	1.1
	309	302	1.0
	139	129	0.9
	11	9	0.8
	565	365	0.6
	Undetectable	500
	Precipitated	330

Rabbit and mouse antibodies

Protein	Expi293 or current system (mg/L)	ExpiCHO (mg/L)	Fold increase
Rabbit antibody	5	120	20
Rabbit antibody	17	178	11
Murine antibody	216	1,412	6.5
	20	86	4.3
	60	240	4.0
	60	240	4.0
	76	196	2.6
	12	18	1.5
	16	17	1.1
	119	48	0.4
	135	47	0.3

Other antibodies

Protein	Expi293 or current system (mg/L)	ExpiCHO (mg/L)	Fold increase
Bi-TE	Western blot	Western blot	25
Bi-specific	28	117	4.2
	26	90	3.5
	126	250	2.0
	NA	763
	NA	660
Secreted	216	1412	6.5

Non-antibody proteins

Protein	Expi293 or current system (mg/L)	ExpiCHO (mg/L)	Fold increase
Non-Ab	15	82	5.5
	Western blot	Western blot	5.0
	164	68	0.4
	NA	206
Fab	85	366	4.3
Fab	10	10	1.0
FC-fusion	5	8	1.7
His Tag HLA	Undetectable	12	12
Viral protein	Western blot	Western blot	3–5
Cytokine	Western blot	Western blot	5–10
FcR	Western blot	Western blot	2–3

ExpiCHO Expression System cost-effectiveness compared to other transient expression systems

	ExpiCHO Expression System	Expi293 Expression System	FreeStyle CHO Expression System
Cost of 1L transfection	$2,174	$1,735	$1,201
Culture volume for equivalent yield	70 mL	100 mL	2,000 mL
Total cost for same amount of protein	$146	$174	$2,403
*Factors in minimum cost of a MaxCyte processing assembly and a portion of the upfront cost of purchasing a MaxCyte electroporation instrument. NOTE: Cost comparison based on list price in USD and protein yields of 1.5 g/L with ExpiCHO Expression System, 1 g/L with Expi293 Expression System, and 0.05 g/L with FreeStyle CHO Expression System.

Choose a system that flexes to fit every phase of your development process

The ExpiCHO Expression System provides multiple protocols to fit your workflow. The system can scale up or down based on your needs for greater throughput early in discovery or higher yields as you focus on selected proteins.

“The ExpiCHO system is so easy it takes less work than making proteins in E. coli. The scalability of the system is fantastic. I can express 30 different mAbs simultaneously, and then pick the ones I like and scale up to grams. The ExpiCHO system is a revolutionary lifesaver for us.” Arjen van den Berg, PhD, Senior Scientist, Cellular and Molecular Medicine, University of California at San Diego

Figure 2. Scalability of the ExpiCHO System. The ExpiCHO System is scalable within 15% of control from 35 mL to 1 L culture volumes.

Figure 3. Transient transfection protocol—7 or 8 steps to protein production. The ExpiCHO system allows you to customize your work with a choice of 3 different protocols depending on your unique research needs, time, and equipment availability.

Fully documented cGMP-banked ExpiCHO-S cells

Fully documented cGMP-banked cell line
Derived from the same Master Seed Bank as standard ExpiCHO-S cells (Cat No. A29127)
Pre-adapted to ExpiCHO Expression Medium
A component of the fully integrated, animal origin-free ExpiCHO Expression System for applications requiring cGMP-banked cells
Flexible commercial licensing options with no royalties

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ExpiCHO cGMP banked cells produce IgG titers equivalent to non-cGMP-banked line

Figure 4. Equivalent rabbit IgG titers are obtained in cGMP-banked ExpiCHO-S cells and non-cGMP ExpiCHO-S cells at 30 mL scale using ExpiCHO-S Expression System.

Easier transition to stable production with ExpiCHO Stable Production Medium

Gibco ExpiCHO Stable Production Medium (SPM) is specifically designed to simplify your cell line development process, facilitating the production stage with minimal effort and a high degree of confidence. This chemically defined, protein-free, animal origin component-free formulation is fully optimized for large-scale fed-batch culture with ExpiCHO-S cells. ExpiCHO SPM supports transition from transient to stable production and offers high titers typically without the need for additional medium optimization.

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Efficiently scale ExpiCHO protein production with Nalgene 5 L Angled-Bottom Shake Flask

Nalgene 5 L Angled-Bottom Shake Flask product photo

Discover Nalgene 5 L Angled-Bottom Shake Flask designed for ExpiCHO expression system scale up. In contrast to other 5 L shake flasks that can impart damaging levels of shear stress on mammalian cells, the unique design of the Nalgene 5 L Angled-Bottom Shake Flask creates a gentle cellular environment allowing for cell growth and expression levels equivalent to those of small-scale shake flasks. The Nalgene 5 L Angled-Bottom Shake Flask was the only 5 L flask tested that enabled such highly efficient scale-up.

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Graph of protein expression in different flasks

Figure 1. ExpiCHO protein expression in Nalgene 5 L Angled-Bottom Shake Flasks compared to alternative 5 L flasks. Protein titer for ExpiCHO cells transfected, resulting in a 2 L final volume in Nalgene 5 L Angled-Bottom flasks compared to other 5 L flasks. Cultures were harvested at Day 14 posttransfection to determine protein titers. Protein titers were >10-fold for the Nalgene Angled-Bottom flasks compared to alternative 5 L flask designs.

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