Anti-PEG antibody ELISA (human IgM specific)

Polyethylene glycol (PEG) chains are often used to modify therapeutic biologic agents in order to prolong the circulating half-life of the modified protein. It has been reported that repeat injections of PEGylated proteins can induce anti-PEG antibodies. This assay employs the sandwich enzyme immunoassay technique. Immobilized PEG is coated onto a 96 well microplate. Calibrator and test samples are pipetted into the appropriate wells. Anti-PEG antibodies present in biological matrices is bound by the immobilized PEG. After washing away any unbound substances, enzyme linked anti IgG or IgM antibody is added to the wells. The plate is washed to remove any unbound antibody-enzyme reagent and a substrate solution is added to the wells for color development. The color development is proportional to the amount of anti-PEG antibodies present in test samples. The color development is stopped and the intensity of the color is measured