Mammalian cell expression system has the most complicated posttranslational modifications closest to the native ones. Posttranslational modifications are required for appropriate biological activity, immunogenicity, and biodistribution. . Modifications such as the authentic glycosylation are essentially only provided by mammalian cells. For this reason, therapeutic and bioactive proteins are usually expressed in mammalian system.
Novoprotein’s mammalian expression system serves you with these advantages:
Our proprietary vectors support high-level expression of proteins in mammalian cells up to 200mg/L.
Two Expression Options:
We offer both transient transfection and stable cell expression
Our serum-free suspension cell culture (HEK293 and CHO) allows for gram scale protein production within one month.
10 transient expression projects can be conducted in parallel.
High Success Rate:
We have successful expressed more than 500 proteins in our mammalian system with yields above 1mg/L.
Timeframe and pricing
- Your gene of interest
- Evaluate and choose the suitable expression system
- Codon Optimization
- Sub-clone into Expression Vectors
- Transient trasfection
- Establish stable cell line
- Expression evaluation and optimization
- Large scale expression
(up to 100L, gram scale)
- SDS Page Gel
- Affinity column
- Ion exchange chromatography
- Hydrophobic chomatography
- Size exclusion chromatography
- Ready to go!
|Gene Cloning and Expression Test|
|Strategy design||Free||1 day|
|Codon optimization||Free||2-3 weeks|
|Subcloning gene of interest into expression vector||$195.00|
|Detect protein expression 24-72 hours post transfection at 12-hour intervals||$500.00||1 week|
|Selection of high-expression stable cell line (optional)||$3,000.00-$5,000.00||2-3 weeks|
|Pilot Scale Expression and Purification|
|1 L HEK293 or CHO cell culture||From $1,500.00||6-12 weeks|
|1mg of protein (soluble, > 80% purity)|
|Large Scale Production|
|Large-scale fermentation up to 500L with 60-80g cell pellet/L||From $2,500.00||4-8 weeks|
|Purification to gram-scale protein|
|Protein production to kilogram scale||Please inquire|
- secreted protein, with two 14kDa subunits
- purify non-covalently linked, no tagged heterodimers
- Clone each subunit to an individual construct
- Co-transfection with two subunit constructs at 1:1 ratio
- Blue Gel affinity: Purify based on binding properties
- Ion exchange (4 exchanges in total) : Purify based on theoretical PI value
- 6 weeks
- 17mg heterodimer protein above 95% purity
1. Purification I – affinity blue gel: cell culture supernatant loadedFig. 1 SDS-PAGE analysis of protein after affinity purification. Targeted protein bound to blue gel column.
4: Elution (reduced);
8: Elution (Non-Reduced)
2. Purification II – cation exchange purification (SP-HP): elution of affinity blue gel desalted and loadedFig. 2 SDS-PAGE analysis of protein after SP-HP ion exchange purification. Targeted protein bound at pH 6.0.
1: sample; 2-3: Flow-through, 4-8: Elutions 1-5
3. Purification III – 1st anion exchange purification (Q-HP): elution of SP-HP desalted and loadedFig. 3 SDS-PAGE analysis of protein after Q-HP ion exchange purification. Targeted protein bound at pH 8.0.
1: sample; 2-5: Flow-through, 6-8: Elutions 1-3
4. Purification IV – 2nd anion exchange purification (Q-HP): elution of 1st Q-HP desalted and loadedFig. 4 SDS-PAGE analysis of protein after Q-HP ion exchange purification. Targeted protein binding improved yet incomplete at pH 8.8.
1: sample; 2: Flow-through, 3-6: Elutions 1-4; 9-10: Elutions (non-reduced)
5. Purification V – 3rd anion exchange purification (Q-HP): elution of 2nd Q-HP desalted and loadedFig. 5 SDS-PAGE analysis of protein after Q-HP ion exchange purification. Targeted protein binding is complete at low temperature.
1: sample; 2: Flow-through, 3-5: Elution (reduced); 9-10: Elution (non-reduced)
6. Final product: Concentration 0.5mg/ml with purity greater than 95%Fig. 6 SDS-PAGE analysis of final product.
1: Reduced; 3: Non-reduced (heterodimer)
For more information about our mammalian expression service, contact us at email@example.com or visit our Service page. In addition, we also offer other protein expression services including bacteria protein expression service, yeast protein expression services, baculovirus expression services. To know how other scientists speak of us, check out our References.
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Our service team keeps tight monitoring and tracking over each step where data are generated.