Insect expression system allows proper refolding, post-translational modification, and oligomerization that are identical to those that occur in mammalian cells. Expression of recombinant proteins is under the control of the strong polyhedrin promoter, allowing expression of up to 50% of the total cell protein. The baculoviral genome is large (130 kb) and can thus accommodate large segments of foreign DNA. Therefore, recombinant proteins can be expressed without size limitations.
Novoprotein’s insect expression system serves you with these advantages:
our system relies on the latest technology in order to ensure high-level expression of both intracellular and secreted recombinant proteins.
various insect cell lines, harvest at different time point and test different MOI to guarantee the highest yield.
our expression capacity reaches up to 30L per batch and 90-100L monthly.
Purity greater than 80% starting from $1500!
Timeframe and pricing
- Your gene of interest
- Evaluate and choose the suitable expression system
- Codon Optimization
- Sub-clone into inset Expression Vector
- Shuttle vector containing viral backbone (bacmid)
- Co-transform E. coli to produce recombinant bacmid
- Prepare recombinant bacmid DNA
- Transfect inset cells & titrate baculovirus
- 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||2-3 weeks|
|Construct of recombinant bacmid||$100.00||1 week|
|Baculovirus titration using plaque assay (titer 10^8-10^9pfu/ml)||$800.00||2 weeks|
|Pilot Scale Expression and Purification|
|Infection of 1L of insect cells (Sf-9, Mimic Sf-9, High Five)||From $1,500.00||3-4 weeks|
|1mg of protein (soluble, > 80% purity)|
|Large Scale Production|
|Large-scale fermentation up to 2-10 liters of insect cell||$600.00/L||3-4 weeks|
|Purification to gram-scale protein|
- Protein difficult to purify
- Two-step purification - Heparin & SP columns
- Step gradient & linear gradient
- refolded bioactive mature peptide above 95% purity
1. Concentration and 1st Purification – His Affinity Column (20ml)
- 6 liters cell culture supernatant concentrated by VIVA200 (COMW 30k) ultrafilter to 1.2L.
- Concentrated solution was purified by 10ml His affinity column. Protein bonded to His column was eluted by step imidazole gradient.
- Elute fraction 3 was pooled for 2nd purification
A: Cell culture supernatant (before dialysis)
B: Cell culture supernatant (after dialysis)
D: Concentrated solution
E: Flow-through of His column
F-H: Elute fractions 1-3
2. 2nd Purification – His HP Column (5ml)
- Protein bound to His HP column was eluted by three step imidazole gradient
- Elute fraction 3 was pooled as final product
F:Tail of fraction 3
3. Final product
For more information about our yeast expression service, contact us at firstname.lastname@example.org or visit our Service page. In addition, we also offer other protein expression services including bacteria protein expression service, yeast expression services,mammalian expression service. 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.