iLite VEGF Assay Ready Cells

$1,240.00

The Eagle Biosciences iLite Assay Ready 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.  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. The iLite VEGF Assay Ready Cells are for research use only.

iLite VEGF Assay Ready Cells

For Research Use Only

The Eagle Biosciences iLite VEGF Assay Ready Cells are intended for testing:

  • VEGF activity
  • NAb against VEGF activity
  • Functional VEGF activity

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.


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

Product Developed and Manufactured by Svar Life Science

Additional Information

Background


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.

Assay Principle


The iLite™ VEGF Assay Ready Cells are engineered cells optimized to express Firefly luciferase under the control of a VEGF responsive promoter. Binding of VEGF to the VEGF receptor 2 (VEGFR2) results in activation of the VEGF regulated Firefly luciferase reporter gene construct. The Firefly luciferase signal can be measured in a luminometer following addition and incubation of luciferase substrate. The Firefly luciferase signal is proportional to the functional activity of VEGF in the sample. In the presence of inhibitory activity against VEGF, the amount of free VEGF is reduced, resulting in a decreased stimulation of Firefly luciferase production. The Firefly luciferase signal is thus inversely proportional to the amount of inhibitory activity against VEGF in a sample. The iLite™ VEGF Assay Ready Cells can therefore be utilized as a highly sensitive assay for quantification of VEGF inhibitor activity in test samples, including human serum.

Assay Quick Guide


Click here for the Assay Summary: VEGF Activity Protocol

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

Frequently Asked Questions


Manual

Product Manual


Publications

References