PathHunter® Aflibercept Bioassay HS Kit
The PathHunter® Aflibercept Bioassay HS Kit is an easy-to-use cell based assay to measure drug potency and detect neutralizing antibodies. This bioassay assesses ligand (e.g. VEGF-165) based activation of KDR/KDR Kinase activity via detection of Dimerization.
Catalog #: 93-0996Y1-00153
Size:
2-Plate
Unit Price: Contact distributor for pricing
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Overview
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Background
VEGF-A is a key protein in forming new blood vessels and increasing vascular permeability. In certain eye diseases, its overproduction causes abnormal vessel growth and fluid leakage, causing vision loss. Aflibercept is a recombinant fusion protein that acts as a decoy receptor, binding VEGF-A and B. By sequestering VEGF-A, aflibercept prevents its interaction with VEGFR on endothelial cells, blocking VEGFR-2 dimerization and the activation of intracellular tyrosine kinase domains. This inhibition of dimerization halts downstream signaling pathways, reducing abnormal angiogenesis, thereby preserving vision.
PathHunter Aflibercept Bioassay Kits models this molecular MOA in a simple, homogenous format, suitable for implementation in characterization, comparability studies, potency testing in QC lot release, stability program, and detection of neutralizing antibodies.
Product Highlights
- Designed for interrogating ligand mediated KDR/KDR dimerization
- Simple, homogenous protocol with results in less than 24 hours
- Significantly better assay performance compared to difficult HUVEC assay
- Suitable for rank ordering, comparability studies, potency measurement, stability studies and neutralizing antibody detection
- Applicable across various stages from characterization and QC lot-release
Assay Principle
The PathHunter Aflibercept bioassay utilizes EFC-based platform and is designed to detect VEGF-A-induced functional homodimerization of the kinase insert domain (KDR), also known as the VEGFR2 receptor. Activation of the KDR receptor through VEGF-A leads to receptor dimerization, which is an essential event in the receptor's signaling cascade. A. The PathHunter KDR/KDR bioassay cells have been engineered to co-express KDR fused to PK, and another KDR construct fused to EA. B. Upon activation by VEGF-A, the VEGFR2 receptors naturally dimerize forcing the two fragments to complement and create an active β-gal enzyme. Active β-gal hydrolyzes its substrate and produces a chemiluminescent signal, indicating receptor activation C. Anti-VEGF molecules like aflibercept, inhibit VEGF-A’s ability to activate the receptors resulting in a dose-dependent reduction of the chemiluminescent signal.
