KRIBIOLISA™ Anti-Darbepoetin (ARANESP™) ELISA
Enzyme Immunoassay for quantitative detection of Anti-Darbepoetin in serum and plasma.
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Introduction:
Erythropoietin (EPO) is a heavily glycosylated protein with a molecular weight of about 30,000 - 34,000 Daltons. Human EPO is a polypeptide consisting of 165 amino acids, containing one O-linked and three Nlinked carbohydrate chains. The recombinant EPO is a good substitute for the native protein for use in an immunoassay. Darbepoetin is a re-engineered form of erythropoietin, contains 5 N-linked oligosaccharide chains and has a molecular weight of 37,100 Daltons and a carbohydrate composition of 51%.It has a 3-fold longer serum half-life compared to epoetin alpha and epoetin beta.
Intended Use:
The KRIBIOLISA™ Anti-Darbepoetin ELISA is used for quantitative detection of Anti-Darbepoetin in 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 D a r b e p o e t i n . 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.
