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:
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 (low titer). 2. P2 generation virus production (high titer). 3. Small volume cell culture and expression test. 4. Protein purification (1-step, affinity purification). 5. SDS-PAGE test. 6. QC and report. Addition: > Gene codon optimization, synthesis and
reconstructed plasmid. |
1. 0.2mg-1mg purified protein. 2. Data and QC report. |
|
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. Midi volume (XXX mL) cell
culture and expression condition test: up to 3 different conditions. 5. Protein purification (1-step,
affinity purification). 6. WB and SDS-PAGE test. 7. QC and report. |
1. 0.5mg-5mg purified protein. 2. Data and QC report. |
|
P110413 Insect-Comfo 9 – 11 Weeks |
1. Condon optimization and gene
synthesis (free of charge). 2. 2 expression vectors will be used
for reconstructed plasmid. 3. 2 cell lines will be used to
test expression. 3. P1 generation virus production
(low titer) and expression test. 4. P2 generation virus production
(high titer) and expression test. 5. 200mL - 1L cell culture. 6. Protein purification with 2 steps, purity polishing to >85%. 7. WB and SDS-PAGE test. 8. QC and report. |
1. 1mg - 10mg purified protein. 2. Data and QC report. |
|
P110402 1L Cell Culture 2 Weeks |
1L insect cell culture and protein
purification for 1 clone. |
Up to 30mg purified protein. |
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. |
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 |
|
Case study 1 - Virus proteinA 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 - Enzyme protein and bioactivity testAn 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. |
Step 1 – P1 generation virus infection and expression test.
Reconstructed bacmid infected both Sf9 and Hi5 cell line. The protein was successfully secreted to culture medium as well as soluble Intracellular expression (result only show Sf9). The intracellular soluble fraction was selected by client for further study. M: protein marker; Φ: non-infected Sf9 cell; 1 and 2: clone 1 and clone 2 both expressed by Sf9 cell; Culture medium: protein expression in culture medium; Soluble protein: soluble expression in intracellular; Insoluble protein: insoluble expression in intracellular. |
Step 2 – P2 generation virus MOI test
Virus was collected and infected Sf9 and Hi5 cells, separately. Different MOIs (a, b, c in the images above) were applied during the test to find out the best MOI. 2 different time points (D-A, D-B) fractions were analyzed. Protein expression level: Sf9 is 230mg/L and Hi5 is 280mg/L. M: protein marker; Φ: non-infected Sf9 cell and Hi5 cell; D-A and D-B: cell culture days; a, b and c: different MOI. |