iLite® VEGF Assay Ready Cells :
- The iLite® VEGF Assay Ready Cells can be used for the quantification of VEGF activity, VEGF inhibitor activity and for determination of neutralizing antibodies against VEGF inhibitors in human serum.
- Content: >250 µL of Assay Ready Cells suspended in DMEM medium with 20% heat inactivated fetal bovine serum (FBS) and 10% dimethyl sulfoxide (DMSO).
- Storage: -80°C, Cells should be used within 30 min of thawing.
- Application notes for the following assays are available:
Product Developed and Manufactured by EuroDiagnostica
For Research Use Only
Vascular Endothelial Growth Factor (VEGF) plays a critical role in both normal vascular growth and pathological angiogenesis and the humanized anti-VEGF antibody bevacizumab (Avastin(TM), Genentech) is used widely as a first-line therapy for colorectal cancer and other neoplastic diseases. Current methods for quantifying the activity of human VEGF, or antibodies that neutralize its activity, are bioassays based on the ability of anti-VEGF antibodies to inhibit the proliferation or migration of primary human umbilical vein endothelial cells (HUVEC) or other cells expressing VEGR receptors, following treatment of the cells with VEGF. Such assays can take several days to perform, are subject to a high degree of variation, and are difficult to validate. Reporter-gene assays based on the establishment of a stable cell line transfected with a luciferase reporter-gene placed under the control of a drug responsive chimeric promoter, provide highly sensitive and reproducible methods for quantifying drug activity.
iLite® VEGF Assay Ready Cells is a human embryonic kidney cell line (HEK-293*; ATCC #CRL-1573) that have been genetically engineered and optimized to respond to VEGF with specific, proportional expression of Firefly Luciferase. Normalization of cell counts and serum matrix effects is obtained by a second reporter gene, a NanoLuc Luciferase reporter gene construct, under control of a constitutive promotor.
* The HEK-293 cell line has been used under a license obtained from AdVec Inc.
Why Choose the iLite® VEGF Assay Ready Cells?
- iLite technology employs the cells’ inherent signaling pathways – a true model of cellular events
- Assay Ready Cells – no culturing required!
- No protein tagging or labeling required
- Highly Sensitive and >15x fold induction
- Allows anti-VEGF activity to be quantified rapidly and in a highly specific manner
- Highly reproducible between batches
- Normalization gene allows for consistent results
- Allows direct comparison between biosimilar and innovator drugs in the same assay
- Adjusts for serum matrix effects and differences in cell number
The Eagle Bio iLite Assay Read Cells are designed for the specific detection of drug potency in serum/plasma as well as neutralizing antibodies. They utilize an assay technique called reporter gene assay ( RPG ) to analyze serum/plasma samples. The reporter genes utilized are encoded with a bioluminescent luciferase (Firefly luciferase). Different levels of luminescence detected from each reporter gene cell indicates different levels of expression.
Key benefits of iLite® Assays:
- Highly specific reporter gene cell lines
- Very sensitive cell line responses (>10 fold inductions)
- Assay Ready Cells – ready-to-use from the freezer, without culturing of cells
- Assays within a workday (typically 4-7 hour assays)
- Normalization gene, which eliminates unwanted matrix effects