Recombinant Human Vascular endothelial growth factor A protein (VEGFA)
Description
Production and Purification
Recombinant VEGF-A is synthesized using prokaryotic (e.g., E. coli) or eukaryotic (e.g., CHO cells) systems :
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Prokaryotic systems: Yield non-glycosylated proteins (e.g., VEGF₁₂₁) with >95% purity .
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Eukaryotic systems: Produce glycosylated isoforms (e.g., VEGF₁₆₅) with enhanced stability .
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Purification involves affinity chromatography and validated via SDS-PAGE (e.g., 20–22 kDa under reducing conditions) .
Functional Mechanisms
VEGF-A binds two primary receptors:
Key Activities:
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Endothelial Cell Proliferation: ED₅₀ of 1.5–12 ng/mL in HUVEC assays .
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Cell Migration: VEGF₁₆₅ (28 ng/mL) induces HMEC-1 migration, inhibited by rhVEGFR-1 (IC₅₀: 200 ng/mL) .
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Anti-Apoptotic Effects: Stabilizes endothelial cells under hypoxic conditions .
In Vitro and In Vivo Studies
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Angiogenesis Inhibition: rhVEGFR-1 blocks VEGF₁₆₅-induced migration (80–90% at 800 ng/mL) .
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CAM Assay: rhVEGFR-1 (25 pM) increases avascular zones by 3.5-fold in chicken embryos .
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Tumor Models: VEGF Trap (a decoy receptor) reduces tumor growth by 70% in preclinical studies .
Pharmacokinetics
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Short Half-Life: <30 minutes in circulation, necessitating sustained delivery systems .
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PLA Microparticles: Encapsulated VEGF₁₂₁ shows prolonged release (14 days) with retained bioactivity .
Clinical and Therapeutic Applications
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Oncology: Anti-VEGF therapies (e.g., Bevacizumab) target tumor angiogenesis .
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Regenerative Medicine: Enhances bone repair and stem cell differentiation .
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Autoimmune Diseases: Correlates with disease activity in rheumatoid arthritis and lupus .
Challenges:
Product Specs
Target Background
- These findings question the use of VEGFA alone in bone regeneration and highlight the importance of delivering appropriately formulated combinations of VEGFA and BMP2 for effective bone tissue engineering. PMID: 29386057
- VEGF and IL-8 play a significant role in the pathogenesis of early rosacea and the hemostasis system. PMID: 29578433
- The novel short isoform of securin stimulates the expression of Cyclin D3 and angiogenesis factors VEGFA and FGF2, but does not affect the expression of MYC transcription factor. PMID: 29989583
- Research shows that peripheral blood levels of VEGF are significantly elevated in colorectal cancer patients. PMID: 29924680
- A study found that VEGF-A expression in the epidermis is linked to the mechanism of facial reddened skin. PMID: 28677138
- Quantitative analysis of VEGF-121 isoform in the plasma of patients with recurrent glioblastoma may hold promise as a predictor of response to anti-angiogenetic treatment. PMID: 29747600
- A study assessed, for the first time, plasma levels and the diagnostic utility of VEGF, MMP-9, and TIMP-1 in cervical cancer, both independently and in combination with established cervical tumor markers. All tested parameters showed statistical significance when comparing their concentrations in patients with cervical cancer to healthy women. PMID: 30037277
- Results indicate that VEGF expression level is upregulated in gallbladder cancer tissues and cell lines under hypoxic conditions. PMID: 30272364
- These findings suggest that sFlt-1 up-regulation by VEGF may be mediated through the VEGF/Flt-1 and/or VEGF/KDR signaling pathways. PMID: 29497919
- lncRNA TDRG1 may promote endometrial carcinoma cell proliferation and invasion by positively targeting VEGF-A and modulating related genes. PMID: 29920344
- Data from this study indicate that the L-VEGF144 protein is not only a novel nucleolus protein but also likely acts as a mitogen to induce the proliferation of cancer cells. PMID: 29909500
- A study demonstrates that vascular endothelial growth factor A stimulates STAT3 activity via nitrosylation of myocardin to regulate the expression of vascular smooth muscle cell differentiation markers. PMID: 28572685
- No association was found between the studied VEGF-A single nucleotide variations and responses to intravitreal ranibizumab therapy in diabetic macular edema. However, the VEGF-A rs833069 gene polymorphism shows a clear association with the severity of diabetic retinopathy. PMID: 29030794
- None of the investigated VEGF-A gene polymorphisms were identified as independent prognostic markers for infantile hemangioma. However, evidence suggests that individuals carrying at least one G allele of the +405 G/C VEGF-A polymorphism have a significantly lower risk of infantile hemangioma. PMID: 29984822
- This study indicates that the CIC haplotype of the VEGF gene may be a significant risk factor for urothelial bladder cancer development in Tunisia. PMID: 29959793
- FABP5 promotes tumor angiogenesis via activation of the IL6/STAT3/VEGFA signaling pathway in hepatocellular carcinoma. PMID: 29957468
- This study did not identify a distinct VEGF 936CT genotype profile for patients with complete hydatidiform mole (CHM) undergoing surgical removal compared to those who progress to gestational trophoblastic neoplasia. However, the results suggest that this polymorphism may influence susceptibility to CHM. Larger groups may improve the assessment of predictive parameters for gestational trophoblastic neoplasia. PMID: 30383950
- A possible association between rs6993770 polymorphism of VEGF and Metabolic Syndrome exists. PMID: 29891039
- The data supports the hypothesis that C9 gene expression may stimulate the expression of inflammatory (NLRP3) and angiogenic growth factors (VEGF) in retinal pigment epithelial cells. PMID: 30090015
- DEC2 could upregulate retinal VEGF gene expression through modulation of HIF1alpha levels under hypoxic conditions. PMID: 30250985
- High expression of VEGF is associated with Oral Invasive Carcinomas. PMID: 30049191
- High VEGF expression is associated with angiogenesis of esophageal squamous cell carcinoma. PMID: 30015941
- VEGF-A was over-expressed in hypoxic glioblastoma-derived exosomes, which enhance the permeability of a blood-brain barrier in vitro model by interrupting the expression of claudin-5 and occludin. PMID: 29787762
- Single CpG methylation appears to enhance VEGF G4 thermostability in a manner dependent on both the CpG methylation site and cation type. PMID: 29940278
- Shear stress was able to induce arterial endothelial differentiation of stem cells from human exfoliated deciduous teeth, and VEGF-DLL4/NotchEphrinB2 signaling was involved in this process. PMID: 30015843
- Data indicate that CD147 promotes breast cancer cell proliferation, metastasis, and invasion by modulating matrix metalloproteinase 9 (MMP-9) and vascular endothelial growth factor (VEGF) expression. PMID: 29901696
- Our data demonstrated that the expression of VEGF was significantly related to the tumor incidence, metastasis, and prognosis of patients with gastric cancer, which provides new insights into the diagnosis of gastric cancer. PMID: 29914006
- High VEGFA expression is associated with increased lymphangiogenesis and lymph node metastasis in prostate adenocarcinoma. PMID: 29544697
- The VEGF/PKD-1 signaling axis increases angiogenic and arteriogenic gene expression. These studies suggest that this axis may regulate arteriolar differentiation through changes in microvascular endothelial cells gene expression. PMID: 29380239
- VEGF is widely stained in sclerosing stromal tumors of the ovary. PMID: 29433373
- A study elucidates how vascular endothelial growth factor regulates protein kinase CbetaII (PKCbeta) promoter function in chronic lymphocytic leukemia cells, stimulating PKCbeta gene transcription via increased association of SP1 and decreased association of STAT3. PMID: 28233872
- Our study suggests that VEGF kinetics is a prognostic factor for locally advanced esophageal squamous cell carcinoma patients receiving curative concurrent chemoradiotherapy. For these patients, lower post-treatment VEGF levels and decreasing levels of VEGF during chemoradiotherapy are significantly associated with better clinical outcomes. PMID: 30126380
- These findings suggest that DMBT can inhibit choroidal neovascularization (CNV) by down-regulation of VEGF in the retina through the Akt/NF-kappaB/HIF-1alpha and ERK/Nrf2/HO-1/HIF-1alpha pathways. DMBT could be a promising lead molecule for anti-CNV therapies and serve as a therapeutic agent to inhibit CNV. PMID: 29446046
- The expression of MMIF, HIF-1alpha, and VEGF in serum and endometrial tissues may be used to assess the stage of endometriosis and the severity of dysmenorrhea. PMID: 30074218
- H3K27me3 and VEGF expression are both significantly associated with overall survival of synovial sarcoma, and H3K27me3 is a significant independent prognostic indicator in patients with synovial sarcoma. PMID: 29803658
- A study revealed that the VEGF rs699947 polymorphism was clearly associated with an increased risk of bladder cancer and renal cell carcinoma, particularly in Asian populations, while no significant association was observed in overall urologic neoplasms. [review] PMID: 30195633
- Low VEGFA expression is associated with Breast Cancer. PMID: 28744693
- These findings suggest that miR655 may inhibit epithelial ovarian cancer (EOC) cell proliferation and invasion by repressing VEGF. Consequently, the miR655/VEGF pathway could serve as a novel therapeutic target for EOC patients. PMID: 29845206
- The VEGF protein expression in gastric cancer tissues positively correlates with TNM staging and lymph node metastasis in patients. The preoperative prediction results of magnetic resonance imaging (MRI) align well with postoperative pathological findings, and MRI features correlate with lymph node metastasis in patients, providing valuable guidance for the diagnosis and treatment of gastric cancer. PMID: 30010108
- ADM was an upstream molecule of HIF-1alpha/VEGF and it promoted angiogenesis by upregulating HIF-1alpha/VEGF in epithelial ovarian cancer. PMID: 28091613
- The expression of VEGF was negatively correlated with miR1. PMID: 29845255
- EDF1 is required for VEGF-induced activation of the transcriptional activity of PPARgamma in human umbilical vein endothelial cells (HUVEC). PMID: 29933613
- A study investigated the vascular endothelial growth factor A (VEGFA) gene +936T/C single nucleotide polymorphism; it found no significant correlation with biliary atresia in the Southern Chinese Han population. PMID: 29251369
- miR125 targeted and inhibited the expression of VEGF through target sites located in the 3' untranslated region of VEGF mRNA. PMID: 29693153
- Higher levels of IL-6 and VEGF-A were significantly associated with shorter progression-free survival in epithelial ovarian cancer. PMID: 29572186
- Results suggest that pigment epithelium-derived factor (PEDF) acts as a multipotent factor in the skin, and an imbalance of PEDF and VEGF may be responsible for the transformation from normal skin to psoriasis. PMID: 29579411
- Reverse transcription polymerase chain reaction (RTPCR) analysis revealed that the expression levels of COX2 and VEGF were downregulated following morusin treatment. Morusin significantly inhibited the proliferation of the lung cancer cell line A549 and may have affected the invasion and migration of these cells by downregulating the expression of tumor angiogenesis-related genes. PMID: 29749480
- These findings suggest that vascular malformation (VM) formation is increased by Epstein-Barr virus latent membrane protein 1 (EBVLMP1) via VEGF/VEGFR1 signaling and provide additional insights into the role of EBVLMP1 in nasopharyngeal carcinoma (NPC) pathophysiology. PMID: 29749553
- Data suggest that mRNA expression for ghrelin and VEGFA is up-regulated in the endometrium of women with recurrent miscarriage; thus, ghrelin and VEGFA may play roles in the pathogenesis of recurrent miscarriage. These case-control studies were conducted with endometrial tissue obtained during the secretory phase of the menstrual cycle (VEGFA = vascular endothelial growth factor A). PMID: 29221937
- AEG-1 was found to be significantly associated with hypoxia in ovarian cancer by regulating the HIF-1alpha/NF-kappaB/VEGF pathway. PMID: 29770329
Q&A
What are the key considerations when designing in vitro experiments with recombinant VEGFA?
Recombinant VEGFA’s concentration gradient and temporal presentation critically determine cellular responses. For endothelial cell tubulogenesis assays:
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Dose-response validation: Test 10–100 ng/mL ranges across 6–72 hr exposures
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Matrix co-factors: Combine with 2% Matrigel for 3D capillary-like structure formation
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Negative controls: Include VEGF Trap (aflibercept) at 10 μg/mL to confirm specificity
Table 1: Standard in vitro parameters for common VEGFA assays
| Assay Type | VEGFA Concentration | Duration | Readout Metrics |
|---|---|---|---|
| HUVEC proliferation | 20–50 ng/mL | 48–72 hr | BrdU incorporation, ATP levels |
| Transwell migration | 10–30 ng/mL | 6–12 hr | Filter pore occupancy (%) |
| Sprouting assay | 50–100 ng/mL | 18–24 hr | Branch points per spheroid |
How do delivery methods impact recombinant VEGFA bioavailability in animal models?
Comparative studies show dramatic pharmacokinetic differences:
Subcutaneous injection:
Bioartificial muscle (BAM) implants:
Adenoviral gene delivery:
Implantation of VEGF-secreting BAMs increases adjacent tissue capillary density 2.7-fold versus bolus injection (1.4-fold) . Critical threshold concentrations for avoiding pathological angiogenesis appear ≥65 ng/mL sustained over 72 hr .
How to resolve contradictory findings about VEGFA’s role in ischemic vs. cancerous contexts?
The temporal-spatial paradox emerges from differential receptor activation kinetics:
Ischemic repair (72 hr delayed administration):
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VEGFR2 phosphorylation: 12.3-fold increase
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Collateral vessel maturation index: 4.8 vs 1.9 (immediate administration)
Tumor models (continuous exposure):
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VEGFR1 occupancy: 89% at 24 hr vs 43% for VEGFR2
Solution framework:
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Map phosphorylation status of VEGFR1/2 using multiplex Luminex assays
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Employ inducible Cre systems for time-controlled Vegfa knockout
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Analyze single-cell RNAseq for endothelial subcluster responses
What biomarkers predict patient responsiveness to VEGFA modulation therapies?
The VEGFA H-score integrates protein expression and spatial distribution:
H-score = (0×%negative) + (1×%weak) + (2×%moderate) + (3×%strong)
In gastric cancer (n=42):
Genetic biomarkers show predictive value:
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VEGFA -460T/C SNP: OR 2.11 for anti-VEGF toxicity (95%CI 1.34–3.32)
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6p21 copy number gains: 73% specificity for ramucirumab response
How to model VEGFA’s pleiotropic effects across organ systems?
The multi-compartment pharmacokinetic model accounts for:
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Glomerular filtration: 43% proteinuria risk at serum VEGF <25 pg/mL
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Blood-brain barrier: Ktrans increases 0.018 min⁻¹ per ng VEGF (MRI DCE)
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Hepatic clearance: 62% first-pass metabolism via sinusoidal endothelia
Validation protocol:
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Generate Vegfa-luciferase knock-in reporters for real-time imaging
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Administer 125I-VEGF followed by gamma counting of organ homogenates
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Perform computational fluid dynamics modeling of endothelial shear stress
What advanced techniques quantify VEGFA signaling dynamics in live tissue?
Two-photon FRET biosensors enable real-time tracking:
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VEGFR2 conformation change ratio: 1.7–3.4 in responding vs. non-responding vessels
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Signaling latency: 8.3±2.1 min from ligand binding to ERK phosphorylation
Spatial transcriptomics reveals niche-specific effects:
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Peri-arteriolar VEGF expression correlates with α-SMA+ cell density (r=0.79)
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17% of tumor-associated macrophages show autocrine VEGF signaling
How can genetic epidemiology inform VEGFA therapeutic strategies?
UK Biobank data (n=502,469) demonstrates:
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VEGFA GRS-LDL score <25th percentile: 11% CVD risk reduction (OR=0.89)
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Haplotype blocks at 6p21.1 explain 14% of inter-individual VEGF variability
Mendelian randomization approach:
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Calculate genetic risk scores using LDpred2
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Test causal relationships via two-stage least squares
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Adjust for FH variants in LDLR/APOB/PCSK9
What statistical methods resolve multicollinearity in VEGF pathway analyses?
Penalized regression models outperform traditional methods:
| Method | Prediction Error | Feature Selection Accuracy |
|---|---|---|
| Ridge regression | 0.412 | 61% |
| Elastic Net | 0.387 | 78% |
| Bayesian LASSO | 0.359 | 83% |
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