Baculovirus Vector Development Service
Creative Biolabs delivers a complete range of services for the scaled-up amplification, concentration, purification, and formulation of baculoviruses vector, aiming to fulfill the requirements of customers worldwide.
Baculovirus Vector Introduction
Baculovirus vector is defined as a gene therapy vector that effectively transduces large non viral DNA packages in both dividing and non dividing cells. Due to its status as an insect pathogen, no pre-existing immunity has been observed in humans, making it a safe alternative to other viral vectors. Gene transfer and expression have been successfully carried out in various cell lines of different species and tissue sources using rod-shaped viruses as vectors. Although the target gene delivery system of rod-shaped viruses has made rapid progress, the specific pathway for rod-shaped viruses to enter mammalian cells is still unclear. Currently, it is widely believed that rod-shaped viruses achieve this through endocytosis, and the surface of rod-shaped viruses displays VSVG, which can significantly enhance their ability to transfer exogenous DNA.
Fig.1 Putative model of internalization of baculoviruses into mammalian cells.1
What Do You Get?
Creative Biolabs provides a comprehensive range of services related to rod-shaped virus vectors, covering the entire field of recombinant protein production and gene therapy applications.
- Customized baculovirus vector construction: Design and synthesis of customized transfer vectors, gene cloning, and production of recombinant baculoviruses encoding genes of interest. This includes multi gene expression vectors used for complex protein assembly.
- High titer baculovirus production: Production of high titer, scalable baculovirus stocks (P0 to P3) in a range of insect cell lines, to produce sufficient virus particles for expression at any scale.
- Recombinant protein expression service: Small to large scale protein expression in insect cells, optimizing infection conditions (MOI, cell density, incubation time) to achieve the best yield and solubility.
- Virus like particle (VLP) production: a professional service for VLP expression and assembly, for vaccine development and structural biology research.
- Using BEVS to produce adeno-associated virus (AAV): a highly scalable platform for producing recombinant AAV vectors for gene therapy applications, utilizing the advantages of insect cell culture to achieve high yields and reduce manufacturing costs.
Typical Project Workflow for Baculovirus Vector Development
Creative Biolabs provides the cell lines Sf21, Sf9 and High Five to produce baculovirus vectors and recombinant proteins for both research and manufacturing uses. We have established high-efficient propagation methods and a range of product testing procedures to ensure the production of high titers of baculovirus vectors free of endogenous viruses and incidental contaminants.
| Step | Description |
|---|---|
| 1. Target Design & Codon Optimization | Analyze the customer's gene of interest (GOI) to optimize for insect/mammalian cell expression. |
| 2. Transfer Plasmid Construction | Clone the optimized GOI into a baculovirus transfer plasmid (e.g., pVL1393) with selectable markers (e.g., ampicillin resistance). |
| 3. Bacmid Generation & Transfection | Transform the transfer plasmid into E. coli to generate a recombinant bacmid (viral genome), then transfect into Sf9 cells for viral production. |
| 4. Viral Amplification & Titering | Amplify the recombinant baculovirus (P1 stock) to generate high-titer P2/P3 stocks, then titer via plaque assay or qPCR. |
| 5. Final QC & Delivery | Test vector integrity, sterility, and expression efficiency before delivery. |
Service Highlights
Mammalian Cell-Compatible Vectors: Incorporate SV40 origin or WPRE element to enhance transgene persistence in HEK293 or CHO cells. A 2024 collaboration with a gene therapy firm resulted in vectors with 60% longer transgene expression in vivo (mouse model) vs. standard baculoviruses.
Multigene Expression Vectors: Design dual-promoter plasmids for co-expression of two proteins. For a customer studying protein-protein interactions, we delivered a vector co-expressing luciferase and its substrate, enabling real-time binding assays.
Comprehensive Post-Translational Modifications
This ensures the production of biologically active and functionally relevant recombinant proteins.
Stringent Quality Control
- Rigorous quality control
- Advanced analytical techniques
- Structural authenticity and functional activity analysis
Proprietary Technologies
- Continuous R&D investment
- Optimized vector and cell line developments
- Advanced purification methodologies
Applications of Baculovirus Vector
- Baculovirus Vector Applied in Vaccination
As vaccine carriers, baculovirus vectors can simultaneously stimulate the humoral and cellular immune responses for the treatment of influenza, malaria and rabies. Creative Biolabs provides numerous recombinant baculoviruses with different promoters and regulatory elements to enhance the expression of foreign proteins and boost the efficacy of vaccination.
- Baculovirus Vector Applied in Tissue Engineering
Baculoviruses can be transduced at extremely high efficiency in primary chondrocytes, bone marrow–derived mesenchymal stem cells (BMSCs), and adipose-derived stem cells (ASCs). By expressing various growth factors, baculovirus gene therapy can be used in tissue engineering and regenerative medicine, for example, to regulate cell differentiation and improve cartilage formation.
- Baculovirus Vector Applied in Cancer treatment
Baculoviruses also can express suicide genes, tumor suppressors or other antitumor genes at high efficiency in specific cancer cells. And we provide the design of inserts with immunepotentiating genes to elicit strong host immunity against malignancies of the bladder, brain, lungs and prostate.
Frequently Asked Questions
Q: Is it safe to use rod-shaped virus vectors in human or animal?
A: Yes, it is. Baculoviruses are classified as biosafety level 1 (BSL-1) preparations by the Centers for Disease Control and Prevention and the World Health Organization in the United States, as they do not replicate or cause disease in mammals, humans, or plants. Research has shown that rod-shaped viruses can be rapidly cleared from mammalian tissues (with a half-life of<24 hours in mice) and do not integrate into the host genome.
Q: Can baculovirus vectors express membrane proteins or toxic proteins?
A: Of course. We optimized the vector and culture conditions for membrane protein expression, including using insect cell lines that support appropriate membrane integration and containing companion genes (such as Hsp70) to enhance solubility. For toxic proteins, we provide inducible promoters (such as the ecdysteroid induction system) that allow controlled expression and prevent cell death during viral amplification. A recent customer successfully expressed 50mg/L of toxic ion channel protein using our induction system.
Q: What is the difference between AcMNPV and BmNPV carriers, and which one should I choose?
A: California Autographa californica MNPV (AcMNPV) and silkworm NPV (BmNPV) are the two most common backbone rod-shaped viruses:
- AcMNPV: Wide host range (infected with Sf9, Sf21) Cells have higher protein expression rates for most proteins, making them highly suitable for research and large-scale production.
- BmNPV: It has specificity for Bombyx mori cells or larvae, allowing live silkworms to economically and efficiently produce protein (e.g. 100-200 milligrams of protein per silkworm).
- Our team can help you choose the appropriate backbone based on your cell type, protein type, and size requirements.
Q: How can I increase protein production if my initial expression is low?
A: Our technical team provides troubleshooting: (1) Optimizing Infection Diversity (MOI) - we recommend using 5-10 PFU/cell for Sf9 cells; (2) Extend harvest time (48-72 hours after Polh et al. late promoter infection); (3) Use fusion tags (such as GST) to improve solubility.
Drop Us a Line Today!
Viral vectors have been utilized in gene therapy to delivery therapeutic genes to target cells through ex vivo or in vivo methods. Creative Biolabs provides one-stop services of the scaled-up amplification, concentration, purification and formulation of baculoviruses to meet the needs of customers worldwide. If you have any questions or have any difficulties, you can contact us by email or send us an inquiry to find a complete solution.
Reference
- Ono C, Okamoto T, Abe T, et al. Baculovirus as a tool for gene delivery and gene therapy. Viruses, 2018, 10(9): 510. https://doi.org/10.3390/v10090510. Distributed under Open Access license CC BY 4.0, without modification.
