Custom Baculovirus Service.pdf

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Custom Baculovirus Service 
 
    
                        
   
    
 
 
 
1. Baculovirus Expression Full Service     
 
Catalog No.:  
BCS-10001 
Timeline:  
3-4 weeks 
Required 
Materials:  
20 µg of recombinant baculovirus transfer plasmid containing gene of interest. 
Deliverables:  
Phase 1: 
3 cell pellets for evaluation of baculovirus to be used; 
Phase 2: 
18 sets of cell pellets (including 6 negative controls) and media for 
evaluation of insect host and expression conditions; 
Phase 3: 
500 ml of high-titer recombinant baculovirus stock solution ready to 
express the protein of interest. 
 
  
Service Description 
 
Project Stage 
Description 
Phase 1  
Generation, Identification, 
and Evaluation of 
Baculovirus Recombinants 
Cotransfection of recombinant baculovirus transfer vector containing 
gene of interest with 3 different linearized Baculovirus DNAs. 
Recombinant clones will be identified and amplified. The customer will 
receive 3 cell pellets derived from the amplification for evaluation. The 
customer will choose the best recombinant baculovirus for the protein 
expression. 
Phase 2  
Expression Optimization 
Amplification of the selected recombinant baculovirus stock to a high-titer 
stock of more than 1 x 108  pfu per ml. The high-titer recombinant 
baculovirus stock will be used to infect three insect cell lines (Sf21, Sf9, 
and T.ni) with or without booster virus and cells are harvested at two 
different time points. The customer will choose the best insect host and 
infection conditions. 
Phase 3  
Amplification of High-titer 
Recombinant Baculovirus 
Stock 
Amplification of the selected recombinant baculovirus and titer 
determination of the final high-titer stock by end-point dilution. The 
customer will receive 500 ml of high-titer recombinant baculovirus stock 
as final product.  
 
 
 
 
 
 
 
 
2. Baculovirus Expression Basic Service 
 
Catalog No.:  
BCS-10002 
Timeline:  
2-3 weeks 
Required 
Materials:  
20 µg of recombinant baculovirus transfer plasmid containing gene of interest. 
Deliverables:  
Phase 1: 
3 cell pellets for evaluation of baculovirus to be used; 
Phase 2: 
30 ml of high-titer recombinant baculovirus stock solution ready to 
express the protein of interest. 
 
     
Service Description 
 
Project Stage 
Description 
Phase 1  
Generation, Identification, 
and Evaluation of 
Baculovirus Recombinants 
Cotransfection of recombinant baculovirus transfer vector containing 
gene of interest with 3 different linearized Baculovirus DNAs. 
Recombinant clones will be identified and amplified. The customer will 
receive 3 cell pellets derived from the amplification for evaluation. The 
customer will choose the best recombinant baculovirus for the protein 
expression  
Phase 2  
Amplification of High-titer 
Recombinant Baculovirus 
Stock 
Amplification of the selected recombinant baculovirus and titer 
determination of the final high-titer stock by end-point dilution. The 
customer will receive 30 ml of high-titer recombinant baculovirus stock 
as final product. 
 
 
 
 
3. Subcloning into Baculovirus Transfer Vector 
 
Catalog No.:  
BCS-10006 
Timeline:  
2-3 weeks 
Required 
Material:  
5 µg of plasmid containing gene of interest.  
Complete vector sequence or map. 
Deliverables:  
20 µg of ready-to-use recombinant baculovirus transfer plasmid containing the 
gene of interest 
 
Service Description 
 
• 
Isolating the gene of interest from a supplied vector and subcloning it into a baculovirus 
transfer vector;  
• Verification of recombinant baculovirus transfer vector by restriction mapping and sequencing 
of junctions;  
• Preparation and purification of recombinant vector suitable for co- transfection with 
baculovirus DNA.  
 
4. Amplification of Recombinant Baculovirus Stock 
 
Catalog No.:  
BCS-10003 
Timeline:  
2 weeks 
Required 
Materials:  
At least 5 ml of recombinant baculovirus stock. 
Deliverables:  
Specified quantity (in ml) of high-titer recombinant baculovirus stock solution 
ready to express the protein of interest. 
 
 
Service Description 
• Amplification of the chosen recombinant baculovirus;  
• Titration of final high-titer stock by end point dilution  
 
 
5. Baculovirus Expression Optimization 
 
 
Catalog No.:  
BCS-10004 
Timeline:  
2 weeks 
Required Materials: At least 5-10 ml of recombinant baculovirus stock. 
Deliverables:  
18 sets of cell pellets and media for evaluation of protein yield. 
 
Service Description 
• Amplification of the chosen recombinant baculovirus to a high-titer stock of more than 1 x 108 
pfu per ml;  
• 
Infection of three insect cell lines (Sf21, Sf9, and T.ni using the high-titer stock and with or 
without booster virus;  
• Cells are harvested at two different time points.  
 
Table: Conditions for Expression Optimization  
 
 
Cell Lines Used 
Sf9, T.ni, Sf21 
Booster Virus 
Presence and Absence 
MOI 
10 
Time Points 
72 hours and 96 hours 
 
 
6. Baculovirus Protein Production  
Catalog No.:  
BCS-10005 
Timeline:  
2 weeks (1 - 5 liters), 
 
4 weeks (5 - 20 liters), 
 
8 weeks (20 liters and above) 
Required 
Materials:  
At least 100 ml of high-titer (>108 pfu) recombinant baculovirus stock for each liter 
of culture.  
Optimized protein expression conditions are determined by previous evaluation 
project. 
Deliverables:  
Cell pellets ready for protein purification or in the case of a secreted recombinant 
protein, the customer will receive the supernatant 
 
Service Description 
• Amplification of recombinant baculovirus stock for protein production;  
• Expression of recombinant protein under optimized conditions.  
 
 
Contact our Custom Service Representative today to schedule 
your project:  
Phone: 1-866-672-4436 (US Toll-free) OR 1-858-457-5423 ext. 805 
Fax: 1-858-362-2030 
E-mail: custom@orbigen.com
European customers please contact BioCarta Europe GmbH: 
Phone:  +49-40- 525703-0 
Fax: +49-40-525703-77 
E-mail: info.europe@biocarta.com