Description
Introduction:
Natalizumab (Tysabri) is a humanized monoclonal antibody against the cell adhesion molecule alpha-4-integrin used in the treatment of multiple sclerosis and Crohns disease. It blocks the adhesion of leukocytes to endothelial cells by blocking the interaction of the 4-integrin subunit of 41 with VCAM-1 and of 47 with mucosal MAdCAM-1. This prevents autoreactive leukocytes from exiting blood vessels and entering target organs to cause inflammation (reducing the ability of inflammatory immune cells to attach to and pass through the cell layers lining the intestines and blood-brain barrier). The drug is believed to work by reducing the ability of inflammatory immune cells to attach to and pass through the cell layers lining the intestines and blood brain barrier. Natalizumab has proven effective in treating the symptoms of both diseases, preventing relapse, vision loss, cognitive decline and significantly improving quality of life in people with multiple sclerosis, as well as increasing rates of remission and preventing relapse in Crohn disease. 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 Natalizumab and can be responsible for allergic reaction, or even anaphylactic shock. This ELISA kit detects Anti-Natalizumab antibodies and may be used for monitoring immunogenicity.
Intended Use:
The KRIBIOLISA Anti-Natalizumab ELISA is used as an analytical tool for quantitative determination of Anti-Natalizumab antibodies in human serum and plasma.
Principle:
The method employs the quantitative sandwich enzyme immunoassay technique. Natalizumab is pre-coated onto microwells. Samples and standards are pipetted into microwells and antibodies to Natalizumab present in the sample are bound by the capture antibody. Then, a HRP (horseradish peroxidase) conjugated Natalizumab is pipetted and incubated. After washing microwells in order to remove any non- specific binding, the ready to use substrate solution (TMB) is added to microwells and color develops proportionally to the amount of Anti-Natalizumab in the sample. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm.
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