Description
Introduction:
Ranibizumab (Lucentis) is a recombinant humanized IgG1 monoclonal antibody fragment that binds to and inhibits vascular endothelial growth factor A (VEGF-A). VEGF is a biochemical signal protein that promotes angiogenesis throughout the body and in the eye. Through binding to VEGF-A, ranibizumab interrupts the interaction of VEGF with its receptors, and thus prevents the subsequent growth of new blood vessels. AntiDrug 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 Ranibizumab and can be responsible for allergic reaction, or even anaphylactic shock. This ELISA kit detects Anti-Ranibizumab antibodies and may be used for monitoring immunogenicity.
Intended Use:
The KRIBIOLISA Anti-Ranibizumab ELISA is used as an analytical tool for quantitative determination of Anti-Ranibizumab antibodies in serum, plasma and cell culture supernatant.
Principle:
The method employs the quantitative sandwich enzyme immunoassay technique. Ranibizumab is pre-coated onto microwells. Samples and standards are pipetted into microwells and antibodies to Ranibizumab present in the sample are bound by the capture antibody. Then, a HRP (horseradish peroxidase) conjugated Ranibizumab 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-Ranibizumab in the sample. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm.
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