Description
Introduction:
Infliximab is a monoclonal antibody which has high specificity for TNF alpha and does not neutralize TNF beta. Infliximab causes programmed cell death of TNF alpha expressing activated T lymphocytes, an important cell type mediating inflammation; hence it is generally assumed that resolution of activated T cells by Infliximab explains its efficacy 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 infliximab and can be responsible for allergic reaction, or even anaphylactic shock.
Intended Use:
The KRIBIOLISA Anti-Infliximab (Remicade) ELISA is used as an analytical tool for quantitative determination of Anti-Infliximab antibodies in serum, plasma and cell culture supernatant. It may be used for monitoring immunogenicity.
Principle:
The method employs the quantitative sandwich enzyme immunoassay technique. Infliximab is pre-coated onto microwells. Samples and standards are pipetted into microwells and antibodies to Infliximab present in the sample are bound by the capture antibody. Then, a HRP (horseradish peroxidase) conjugated Infliximab 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-Infliximab in the sample. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm.
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