Description
Introduction:
Etanercept is a biopharmaceutical that treats autoimmune diseases by interfering with tumor necrosis factor (TNF, a soluble inflammatory cytokine) by acting as a TNF inhibitor. It has U.S. F.D.A. approval to treat rheumatoid arthritis, juvenile rheumatoid arthritis and psoriatic arthritis, plaque psoriasis and ankylosing spondylitis. TNF-alpha is the master regulator of the inflammatory (immune) response in many organ systems. Autoimmune diseases are caused by an overactive immune response. Etanercept has the potential to treat these diseases by inhibiting TNF-alpha. 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 Etanercept and can be responsible for allergic reaction, or even anaphylactic shock. This ELISA kit detects Anti-Etanercept antibodies and may be used for monitoring immunogenicity.
Intended Use:
The KRIBIOLISA Anti-Etanercept ELISA is used as an analytical tool for quantitative determination of Anti-Etanercept antibodies in human serum and plasma.
Principle:
The method employs the quantitative sandwich enzyme immunoassay technique. Etanercept is pre-coated onto microwells. Samples and standards are pipetted into microwells and antibodies to Etanercept present in the sample are bound by the capture antibody. Then, a HRP (horseradish peroxidase) conjugated Etanercept 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-Etanercept in the sample. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm.
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