VEGF Human, Baculovirus
Description
Key Steps:
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Baculovirus Construction: VEGF gene insertion into baculoviral DNA (e.g., via transposition) .
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Insect Cell Infection: High-titer baculovirus stocks infect Sf9/Sf21 cells at optimized MOI (multiplicity of infection) .
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Protein Harvest: VEGF is secreted into culture supernatant or retained intracellularly .
Advantages of BEVS:
Angiogenesis and Disease Models
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Endothelial Cell Studies: VEGF₁₆₅ induces proliferation (EC₅₀: 1–6 ng/mL) and migration in HUVEC cells .
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Receptor Internalization: Baculovirus-delivered VEGF activates VEGFR2/Neuropilin-1 complexes, tracked via Rab GTPase vesicle markers .
Therapeutic Development
| Application | Example | Source |
|---|---|---|
| Cancer Therapy | Anti-angiogenic drug screening | |
| Vaccines | BEVS-produced HPV VLPs (Cervarix®) | |
| Gene Therapy | BacMam vectors for in vivo VEGF delivery |
Comparative Analysis with Other Expression Systems
| Parameter | BEVS | Mammalian Systems |
|---|---|---|
| Cost | Low (serum-free media) | High (FBS-dependent) |
| Yield | 10–100 mg/L | 1–10 mg/L |
| Glycosylation | Insect-type (paucimannose) | Human-type (complex) |
| Time | 3–4 days post-infection | 7–14 days |
BEVS excels in producing difficult-to-express proteins but requires glycoengineering for human-like glycosylation .
Research Advancements and Limitations
Enhancements:
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Surface Display: VSV-G or gp64 fusion improves mammalian cell transduction .
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Dual Promoters: SV40 and CMV promoters enable stable expression in hybrid BacMam systems .
Challenges:
Product Specs
APMAEGGGQN HHEVVKFMDV YQRSYCHPIE TLVDIFQEYP DEIEYIFKPS CVPLMRCGGC CNDEGLECVP TEESNITMQI MRIKPHQGQH IGEMSFLQHN KCECRPKKDR ARQENPCGPC SERRKHLFVQ DPQTCKCSCK NTDSRCKARQ LELNERTCRC DKPRRHHHHH H.
Product Science Overview
The VEGF family consists of several proteins, including VEGF-A, VEGF-B, VEGF-C, VEGF-D, and Placental Growth Factor (PlGF). Among these, VEGF-A is the most studied and is often referred to simply as VEGF. VEGF-A binds to two primary receptors, VEGFR-1 (Flt-1) and VEGFR-2 (KDR/Flk-1), which mediate its angiogenic effects .
Human recombinant VEGF is produced using various expression systems, including Escherichia coli, yeast, and baculovirus-infected insect cells. The baculovirus expression system is particularly advantageous for producing complex proteins with post-translational modifications similar to those in mammalian cells .
The baculovirus expression system utilizes insect cells, such as Sf9 or Sf21, to produce recombinant proteins. This system is highly efficient and capable of producing large quantities of protein. The recombinant human VEGF produced in this system is often tagged with a histidine tag to facilitate purification using nickel affinity chromatography .
VEGF is a key player in promoting angiogenesis and vascular permeability. It is extensively studied for its role in cancer, where it promotes tumor growth and metastasis by enhancing blood supply to the tumor. Anti-VEGF therapies, such as monoclonal antibodies and small molecule inhibitors, are used in the treatment of various cancers and other diseases characterized by abnormal angiogenesis .
