Description
Introduction:
Bevacizumab is a recombinant Human IgG kappa monoclonal antibody specific for all human vascular endothelial growth factor A (VEGF-A) isoforms. An angiogenesis inhibitor, it acts by selectively binding circulating VEGF, thereby inhibiting the binding of VEGF to its cell surface receptors. This inhibition leads to a reduction in microvascular growth of tumor blood vessels and thus limits the blood supply to tumor tissues. These effects also lower tissue interstitial pressure, increase vascular permeability, may increase delivery of chemotherapeutic agents, and favor apoptosis of tumor endothelial cells. Additionally, Bevacizumab binds to and neutralizes all human VEGF-A isoforms an bioactive proteolytic fragments, but not mouse or rat VEGF. Anti-Drug Antibodies (ADA) may induce unwanted side effects in biopharmaceuticals. Hence, ADA has been subjected to increase in scrutiny by the regulatory authorities using immunogenicity safety studies. ADA has been observed in pre-clinical and clinical studies, resulting in significant changes in toxicology, pharmacokinetics and efficacy. These effects result from the generation of drug-induced (neutralizing) autoantibodies against Bevacizumab and can be responsible for allergic reaction, or even anaphylactic shock. This ELISA kit detects Anti-Bevacizumab antibodies and may be used for monitoring immunogenicity.
Intended Use:
The KRIBIOLISA Anti-Bevacizumab ELISA is used as an analytical tool for quantitative determination of Anti-Bevacizumab antibodies in serum, plasma and cell culture supernatant.
Principle:
The method employs the quantitative sandwich enzyme immunoassay technique. Bevacizumab is pre-coated onto microwells. Samples and standards are pipetted into microwells and antibodies to Bevacizumab present in the sample are bound by the capture antibody. Then, a HRP (horseradish peroxidase) conjugated Bevacizumab is pipetted and incubated. After washing microwells in order to remove any nonspecific binding, the ready to use substrate solution (TMB) is added to microwells and color develops proportionally to the amount of Anti-Bevacizumab in the sample. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm.
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