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Baculovirus Expression System

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BacuVance™ Baculovirus Expression System


The baculovirus expression vector system (BEVS) is universally recognized as a powerful and versatile system for production of high quality proteins. Using this technology, Baculovirus DNA is used to carry a foreign gene into suitable, insect host cells, which then translate the foreign gene to make recombinant protein. There are several advantages of insect cells over E. coli such as improved solubility, ability to incorporate post-translational modifications, and higher yields for secreted proteins.

GenScript's BacuVance™ Baculovirus/Insect Cell Expression Platform was developed by our in-house team of scientists for virus production and expression of recombinant proteins from baculovirus-infected insect cells in flexible scale. In addition, as the largest gene synthesis supplier in the U.S., GenScript can provide you with free advanced codon optimization specific to Sf9, Sf21, Hi-5, and other insect cell lines, to further enhance the expression of your protein. For customers who wish to establish the best possible protocol for their project and then scale up, GenScript recommends leveraging our sophisticated BacuVance™ Advanced Package to explore different conditions for the expression of target proteins.

Advantages

  • Proprietary Technologies: With BacuVance™ Baculovirus/Insect Cell Expression Technology, we can establish the best possible protocol for your scale-up projects.

  • Superior Baculovirus Preparation: GenScript's recombinant baculovirus preparation services provide guaranteed quality-consistent batch-to-batch virus.

  • Large-scale protein production: Current capacity up to 100 liters per project.

  • Streamlined Services: GenScript can take your project directly from gene synthesis, to downstream activities including recombinant protein expression, stable cell line generation, antibody production and assay development.


  • Customized Package
  • BacuVance™ Standard
    Package
  • Baculovirus Preparation
  • Case Studies

Customized Package

For customers who wish to establish the best possible protocol for their project and then scale up, GenScript recommends using its sophisticated, customizable, in-house platform to explore different conditions for the expression of target proteins.

Milestones Specifications Deliverables Timeline1
1. Subcloning
  • Subcloning into one or more expression vectors
Construct(s) report
2 weeks
2. Viral generation
  • Generation of recombinant Bacmid DNA
  • Transfection of insect cell with recombinant Bacmid DNA
  • Generation of P1 stock (Low titer), P2 stock (High titer), and determination of virus titer by Quantitative-PCR
  • Verify the protein expression for P1 and P2 generation by western blot with anti-His/GST or other antibody.
5 ml, >107 pfu/ml virus stock
2–3 weeks
3. Pilot expression and purification evaluation
  • Infection of insect cells with P2 stock
  • One step affinity purification
    SDS-PAGE gel and Western blot
Expression data
1.5 weeks
4. 1 L expression and purification
  • 1 L insect cell expression and purification
  • One-step purification
  • QC by SDS-PAGE, Western blot2, and MALDI-TOF3
Purified proteins
QC data
1–2 weeks

1Listed is the typical turnaround time at GenScript. Please note that additional requirements may incur additional steps, and likewise longer processing times.

2FREE Western QC data is available for tagged proteins that can be detected by the anti-His or anti-GST antibody. For other tagged proteins, additional charges may apply.

3Other QC data, such as MALDI-TOF, N-terminal sequencing and LC-MS are available for an additional charge.

BacuVance™ Standard Package

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 Timeline1
1. Subcloning
  • Subcloning into one expression vector
Construct(s) report
2 weeks
2. Recombinant viral 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
4. Insect cell expression and purification
  • 1 L insect cell expression and purification
  • One-step purification
  • QC by SDS-PAGE, Western blot2, and MALDI-TOF3
Purified proteins
QC data
1–2 weeks

1 Listed is the typical turnaround time at GenScript. Please note that additional requirements may incur additional steps, and likewise longer processing time.

2 FREE Western QC data is available for tagged proteins that can be detected by the anti-His or anti-GST antibody. For other tagged proteins, additional charges may apply.

3 Optional MALDI-TOF data is available with additional charge.

Protein Insect

Baculovirus Preparation Service

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
Construct(s) report
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
Recombinant Baculovirus Generation
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.

Baculovirus Expression System
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.

Baculovirus Protein Purification
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

Quotations and Ordering

  • To request a quotation, please fill in the Quotation Form and send it to us by email or fax. Download the Quotation Form in following list:
    • For Baculovirus Protein Expression Service, please download the
    • For Baculovirus Preparation Service, please download the
  • To order, please send the completed Quotation Form 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, 860 Centennial Avenue., 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.