Description
Introduction:
Omalizumab, sold under the trade name Xolair, is a medication originally designed to reduce sensitivity to allergens. It has been used to try to control severe allergic asthma, which does not respond to high doses of corticosteroids and less widely for chronic spontaneous urticaria. Omalizumab is a recombinant DNA-derived humanized IgG1k monoclonal antibody that specifically binds to free human immunoglobulin E (IgE) in the blood and interstitial fluid and to membrane-bound form of IgE (mIgE) on the surface of mIgE-expressing B lymphocytes. Unlike an ordinary anti-IgE antibody, it does not bind to IgE that is already bound by the high affinity IgE receptor (Fc epsilon RI) on the surface of mast cells, basophils, and antigen-presenting dendritic cells. 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 Omalizumab and can be responsible for allergic reaction, or even anaphylactic shock. This ELISA kit detects Anti-Omalizumab antibodies and may be used for monitoring immunogenicity.
Intended Use:
The KRIBIOLISA Anti-Omalizumab ELISA is used as an analytical tool for quantitative determination of Anti-Omalizumab antibodies in serum, plasma and cell culture supernatant.
Principle:
The method employs the quantitative sandwich enzyme immunoassay technique. Omalizumab is pre-coated onto microwells. Samples and standards are pipetted into microwells and antibodies to Omalizumab present in the sample are bound by the capture antibody. Then, a HRP (horseradish peroxidase) conjugated Omalizumab 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-Omalizumab in the sample. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm.
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