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. 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 productivity of your protein. For customers who wish to establish the best possible protocol for their project and then scale up, GenScript recommends customers to leverage 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.

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

BacuVance™ Advanced Package For customers who wish to establish the best possible protocol for their project and then scale up, GenScript recommends leverage the sophisticated BacuVance™ platform to explore different conditions for the expression of the target proteins. Milestones Specifications Deliverables Timeline1 1. Subcloning Subcloning into three 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), P3 determine pfu 5 ml, >107 pfu/ml virus stock 3–4 weeks 3. Expression 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 Expression data 1.5 weeks 4. 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.

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.

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 3 –5 weeks

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. 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. 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.

	Email: protein@genscript.com
	Phone: 1-877-436-7274 (Toll-Free) 1-732-885-9188
	Fax: 1-732-210-0262 1-732-885-5878