Introduction:
Rituximab is a monoclonal antibody which has high specificity for CD20 and it is used to treat certain autoimmune diseases and types of cancer. It is also used for non-Hodgkin lymphoma, chronic lymphocytic leukemia, rheumatoid arthritis, and granulomatosis with polyangiitis, idiopathic thrombocytopenic purpura, pemphigus vulgaris, myasthenia gravis and Epstein – Barr virus-positive mucocutaneous ulcers. It is given by slow injection into a vein. 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 Rituximab and can be responsible for allergic reaction, or even anaphylactic shock. This ELISA kit detects Anti-Rituximab antibodies and may be used for monitoring immunogenicity.
Intended Use:
The KRIBIOLISA Anti-Rituximab (Rituxan) ELISA is used as an analytical tool for quantitative determination of Anti-Rituximab antibodies in human serum and plasma.
Principle:
The method employs the quantitative sandwich enzyme immunoassay technique. Rituximab is pre-coated onto microwells. Samples and standards are pipetted into microwells and antibodies to Rituximab present in the sample are bound by the capture antibody. Then, a HRP (horseradish peroxidase) conjugated Rituximab 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-Rituximab in the sample. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm.
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