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Baculovirus-Insect Cell Expression System (BICES) has been widely used for the production of recombinant proteins. It is one of the most powerful, robust, and versatile eukaryotic expression systems available. the BICES offers multiple advantages for protein production in a variety of applications, such as:

• expression of large cDNA fragment;
• expression of cell signaling proteins, cytokines,  kinase and membrane proteins; 
• virus vaccine development;
• synthesis of virus-like particles (VLP); 
• or to express the protein which has failed in mammalian expression system.​

Service highlight:
1. Team members have average 10 years’ experience in Baculovirus-insect expression system, 99% success rate.
2. Multiple expression cell lines and vectors available.
3. Up to 200L fermentation volume, possible reach 100g protein production.

Service and Timeline	Service Description	Deliverables	Price
P110411 Insect-Ready 5 - 7 Weeks	1. P1 generation virus production. 2. 80mL cell culture. 3. Expression condition test: up to 3 different conditions. 4. Protein purification (1-step affinity purification). 5. SDS-PAGE test. 6. QC and report.	1. 0.3mg-1mg purified protein. 2. Data and QC report.	Inquiry​
P110412 Insect-StepUp 9 – 11 Weeks	1. Condon optimization and gene synthesis (1000bp free). 2. P1 generation virus production (low titer). 3. P2 generation virus production (high titer). 4. 200mL cell culture. 5. Expression condition test: up to 6 different conditions. 6. Protein purification (1-step affinity purification). 7. WB and SDS-PAGE test. 8. QC and report.	1. 0.5mg-5mg purified protein. 2. Data and QC report.	Inquiry​
P110413 Insect-Comfo 9 – 11 Weeks	1. Condon optimization and gene synthesis (free of charge). 2. 2 expression vectors will be tested. 3. P1 generation virus production (low titer). 4. P2 generation virus production (high titer). 5. 200mL - 1L cell culture. 6. Protein purification with 2 steps, purity polishing to >90%. 7. WB and SDS-PAGE test. 8. QC and report. 9. One time video meeting.	1. 1mg - 10mg purified protein. 2. Data and QC report.	Inquiry​
P110402 1L Cell Culture 2 Weeks	1L fermentation and protein purification for 1 clone.	Up to 30mg purified protein.	Inquiry​

 

Optional Service 1. Tag remove 2. 5L~200L fermentation 3. Western Blot for protein confirmation 4. Mass Spectrometry More information: Please Email to info@bio-bench.com if you need special service for Baculovirus-insect Expression System.

List of Vectors	Commercial vectors and Bio Bench improved vectors. Please inquire for the up-to-date list.
List of Expression Strain	Sf9, Sf21, Hi5. Please inquire for the up-to-date list.

Comparison of different types of service

Service type Content	Insect cell Ready	Insect cell StepUp	Insect cell Comfo
Codon optimization	-	✓	✓
Gene synthesis	-	✓	✓
Multiple vector or multiple cell line test	-	-	✓
Expression temperature test: HighT °C	✓	✓	✓
P1 generation virus	✓	✓	✓
P2 generation virus	-	✓	✓
Expression duration test (time points)	1	2	2
Fermentation volume	80mL	200mL	200mL – 1L
Affinity purification	✓	✓	✓
2nd purification: purity enhancement	-	-	✓
SDS-PAGE analysis	✓	✓	✓
WB analysis	-	✓	✓
Data, QC and report	✓	✓	✓
Video meeting support	-	-	✓
Purified protein	0.3mg – 2mg	0.5mg – 5mg	1mg – 10mg
	Inquiry​	Inquiry​	Inquiry​

 

Advantages of Baculovirus-Insect Protein Expression System

1. Large gene capacity: ability to carry large gene fragment and express the protein, protein at 90kDa – 250kDa is recommended to be expressed by insect cell;
   
2. Multiple genes expression capacity: A high capacity for multiple genes or a large insert, because of the huge and flexible viral genome (130 kb);
   
3. Proper post-translational modification (PTM): insect cells are higher eukaryotes and able to modify the protein, particularly the glycosylation;
   
4. High biosafety: baculovirus has strict species specificity and naturally does not infect human;
   
5. High expression efficiency: high yield boosted by the strong promoters (polyhedron, p10, etc.), as well as high titers up to 10^8 pfu in late-stage infected cells enable large-scale protein expression;
   
6. High scale expression volume: Easy to amplify and can produce recombinant proteins in large scale, up to XXX L.

Case Studies - Baculovirus-Insect Protein Expression System

Case study 1 - Virus protein

A virus protein was successfully expressed by Sf9 cell line at 60mg/L. After purification, protein purity reaches 95%.
M: protein marker; R: reduced protein; N-R: non-reduced protein.

Case study 2 - Cell signaling pathway protein

A cell signaling path protein failed to be expressed in both CHO and 293 cell lines. After testing in insect cell lines, the protein was successfully expressed by both Sf9 and Hi5 (result only show Sf9).
M: protein marker; Φ: empty Sf9 cell; 1 and 2: sample 1 and sample 2 both expressed by Sf9 cell.

Case study 3 - Enzyme protein and bioactivity test

An enzyme protein was successfully expressed by Sf9 cell line. After purification, protein purity reaches 90%. Then enzyme activity was tested by reagent kit and the signal was read at OD520. The enzyme activity is 420U/mg after testing.
Fig. A - M: protein marker; 1: Elution by 250 mM imidazole; 2: Elution by 60 mM imidazole.
Fig. B – Standard curve of enzyme activity, R^2=0.9982.