Baculovirus Expression System
The Baculovirus Expression Vector System (BEVS) is universally recognized as a powerful, versatile means of producing large quantities of recombinant proteins. Its ease of use, high expression level, and ability to incorporate post-translational modifications make it a valuable part of any protein expression arsenal. The GenScript baculovirus expression service package includes both virus production and flexible-scale protein production. GenScript has pioneered methods for the secretion of recombinant proteins from baculovirus-infected insect cells. Our services are suitable for academic institutions and both the pharmaceutical and biotechnology industries. Our current capacity allows to produce proteins at large quantities, up to 500 liters per project.
GenScript protein expression specialists work closely with clients to clone and express full-length recombinant proteins from full-length cDNAs, EST sequence information, and genomic sequences, providing service and expertise at all stages of protein expression and production.
In addition, as the largest gene synthesis supplier in US, GenScript can provide you with free advanced codon optimization specific to Sf9, Sf21, Hi-5 and other insect cell lines, to further enhance the productivity of your protein. Or use our free rare codon analysis tool to see if your gene sequence is suboptimal, and explore means to boost your target protein expression. Learn more
- BacuVanceTM
Optimization
Package
- Standard Package
- Baculovirus Preparation
- Case Studies
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BacuVanceTM Optimization Protein Expression Package (SC1347)
For customers who wish to establish the best possible protocol for their project and then scale up, GenScript recommends leverage the sophisticated BacuVanceTM platform to explore different conditions for the expression of the target proteins.
| Milestones |
Specifications |
Deliverables |
Timeline* |
| 1. Subcloning |
- Subcloning into three expression vectors
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Construct(s) report
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2 weeks |
| 2. Virus generation |
- Generation of recombinant Bacmid DNA
- Transfection of insect cell with recombinant Bacmid DNA
- Generation of P1 stock (Low titer), P2 stock (High titer), P3 determine pfu
| 5 ml, >107 pfu/ml virus stock |
3–4 weeks |
| 3. Expresion optimization and pilot protein production |
- Infection of insect cells with P3 stock
- Expression optimization and detection:
MOI, time course, cell lines
- SDS-PAGE gel and Western blot
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Expression data |
1.5 weeks |
| 4. Expression and purification |
- 1 L insect cell expression and purification
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One-step purification
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Purified proteins
QC data |
1–2 weeks |
*Listed is the typical turnaround time at GenScript. Please note that additional requirements may incur additional steps, and likewise longer processing time.
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Standard Protein Expression Package (SC1258)
If customers already have established protocols, GenScript offers a standard baculovirus protein expression package. All of the following steps are executed according to the customer's requirements.
| Milestones |
Specifications |
Deliverables |
Timeline* |
| 1. Subcloning |
- Subcloning into one expression vector
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Construct(s) report
|
2 weeks |
| 2. Recombinant virus generation |
- Generation of recombinant Bacmid DNA
- Transfection of insect cell with recombinant Bacmid DNA
- Generation of P1 stock (Low titer), P2 stock (High titer), determine titer
|
5 ml, >107 pfu/ml virus stock |
2–3 weeks |
| 3. Protein expression evaluation |
- Infection of insect cells with P2 stock
- SDS-PAGE gel and Western blot
Expression data |
1.5 weeks |
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| 4. Insect cell expression and purification |
- 1 L insect cell expression and purification
- One-step purification
Purified proteins
QC data |
1–2 weeks |
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*Listed is the typical turnaround time at GenScript. Please note that additional requirements may incur additional steps, and likewise longer processing time.
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Baculovirus Preparation Service (SC1261)
GenScript's recombinant baculovirus preparation services provide guaranteed consistent batch-to-batch virus. We provide titered, highly infectious virus stocks ready-to-use, and detailed storage protocols.
| Milestones |
Specifications |
Deliverables |
Timeline |
| 1. Subcloning |
- Subcloning into one expression vector
|
Expression vector with insert |
2 weeks |
| 2. Virus generation |
- Generation of recombinant Bacmid DNA
- Transfection of insect cell with recombinant Bacmid DNA
- Generation of P1 stock (Low titer), P2 stock (High titer), determine titer
- Protein expression verification
|
Virus stock |
2–3 weeks |
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1. Project evaluation and high secretory expression level (6 mg/L)
Customer’s initial request was to synthesize the gene in question and to clone it into our BV vector for intracellular expression. The expected yield was 0.1–1 mg of tagged protein with over 70% purity. Based on a thorough sequence analysis, we designed a new strategy, which was approved by the customer, to use the protein's native signal peptide to drive secretion. In the end, we obtained 3 mg of protein with a purity level over 80% from 0.5 L SF9 CM. The protein's identity was further confirmed by MALDI-TOF.
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Fig. SDS-PAGE Analysis of target protein purified by SP column
Lane 1–8: Eluted fractions with 1 M NaCl in 20 mM PB, pH 5.0 |
2. Tag-free protein purification
After protein expression, the conditioned medium was dialyzed against buffer, and loaded onto SP column. The predicted size of the target protein was 78KDa. The SDS-PAGE results showed that proteins of the same size as the target had been obtained in lanes 12, 13, 14.
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Fig. SDS-PAGE Analysis of target protein purified by SP column
Lane 1: The condition medium
Lane 2–14: Eluted fractions with gradient concentration of NaCl from 0 to 500 mM in 20 mM PB, pH 5.0 |
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Quotations and Ordering:
 To request a quotation, please download and complete the Quotation/Order Form, and send it to us by email or fax.
To order, please download and complete the Quotation/Order Form and send it to us by email or fax with a formal PO (Purchase Order) or credit card information. You can also submit PO/credit card information by phone or via our secure online messaging system.
If submitting samples, please mail them together with a hard copy of the completed Quotation/Order Form to: Protein Services, GenScript, 120 Centennial Ave., Piscataway, NJ 08854, US.
For questions, or to inquire about the status of your order contact us by email, phone, fax or via our secure online messaging system.
Our customer service representatives are available 24 hours a day, Monday through Friday to assist you.
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Email: protein@genscript.com |
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Phone: 1-877-436-7274 (Toll-Free) 1-732-885-9188 |
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Fax: 1-732-210-0262 1-732-885-5878 |
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