Introduction:
Denosumab (trade names Prolia and Xgeva) is a human monoclonal antibody for the treatment of osteoporosis, treatment-induced bone loss, metastases to bone and giant cell tumor of bone.Denosumab is contraindicated in people with low blood calcium levels.The most common side effects are joint and muscle pain in the arms or legs.Denosumab is a RANKL inhibitor which works by preventing the development of osteoclasts which are cells that break down bone (bone resorption). 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 Denosumab and can be responsible for allergic reaction, or even anaphylactic shock. This ELISA kit detects Anti-Denosumab antibodies and may be used for monitoring immunogenicity.
Intended Use:
The KRIBIOLISA Anti-Denosumab ELISA is used as an analytical tool for quantitative determination of Anti-Denosumab antibodies in serum, plasma and cell culture supernatant.
Principle:
The method employs the quantitative sandwich enzyme immunoassay technique. Denosumab is pre-coated onto microwells. Samples and standards are pipetted into microwells and antibodies to Denosumab present in the sample are bound by the capture antibody. Then, a HRP (horseradish peroxidase) conjugated Denosumab is pipetted and incubated. After washing microwells in order to remove any nonspecific binding, the ready to use substrate solution (TMB) is added to microwells and color develops proportionally to the amount of Anti-Denosumab in the sample. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm.
