Description
Introduction:
Tocilizumab, also known as atlizumab, is an immunosuppressive drug, mainly for the treatment of rheumatoid arthritis (RA) and systemic juvenile idiopathic arthritis, a severe form of arthritis in children. It is a humanized monoclonal antibody against the interleukin-6 receptor (IL-6R). IL-6 is a pleiotropic cytokine with diverse physiological actions, regulating the immune response, inflammation, bone metabolism, and hematopoiesis. It is implicated in the pathogenesis of many diseases, such as autoimmune diseases, multiple myeloma and prostate cancer. IL-6 forms a complex by binding to IL-6 R on the cell membrane, which then combines with gp130, which also resides on the cell membrane, forming a homodimer and giving rise to intracellular signal transduction. Tocilizumab inhibits the induction of biological activity due to IL-6 in cells that have expressed both membrane-bound IL-6R and gp130 molecules, and also inhibits the induction of biological activity due to IL-6/IL-6R complex formation in cells that express gp130 alone. It also exhibits effective blocking of IL-6 signal transduction due to its capacity to dissociate IL-6/IL-6R complexes that have already formed. 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 Trastuzumab and can be responsible for allergic reaction, or even anaphylactic shock. This ELISA kit detects Anti-Tocilizumab antibodies and may be used for monitoring immunogenicity.
Intended Use:
The KRIBIOLISA Anti-Tocilizumab ELISA is used as an analytical tool for quantitative determination of antibodies to Tocilizumab in serum, plasma and cell culture supernatant.
Principle:
The method employs the quantitative sandwich enzyme immunoassay technique. Tocilizumab is pre-coated onto microwells. Samples and standards are pipetted into microwells and antibodies to Tocilizumab present in the sample are bound by the capture antibody. Then, a HRP (horseradish peroxidase) conjugated Tocilizumab 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 antibodies to Tocilizumab present in the sample. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm.
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