Description
Introduction:
Darbepoetin alfa is a re-engineered form of erythropoietin that binds to the erythropoietin receptor on erythroid progenitor cells, stimulating RBC production and differentiation. It stimulates erythropoiesis (increases red blood cell levels) by the same mechanism as rHuEpo (binding and activating the Epo receptor) and is used to treat anemia, commonly associated with chronic kidney failure and cancer chemotherapy. 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-Darbepoetin Alfa antibodies and may be used for monitoring immunogenicity.
Intended Use:
The KRIBIOLISA Anti-Darbepoetin ELISA is used for quantitative detection of Anti-Darbepoetin antibodies in human serum and plasma.
Principle:
The method employs enzyme linked immunoassay technique. Standard or samples are pipetted into microwells pre-coated with Darbepoetin and A nti- Darbepoetin present in the sample and standards are bound by Darbapoetin. In the second step, HRP conjugate is pipetted and incubated. Free HRP conjugate will be removed by washing. Addition of TMB substrate will develop blue color and and reaction is stopped by addition of stop solution. Colour of the solution turns yellow. Intensity of yellow colour in wells is directly proportional to the concentration of Anti-Darbepoetin present in Standard or sample. Absorbance is measured at 450 nm.
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