Information

The PEGylated Protein ELISA kit is a competitive assay specific to the backbone of poly(ethylene glycol) (PEG) and has been validated for use with a wide variety of PEG molecules including linear and branched forms as well as free and conjugated forms. As expected, the assay is more sensitive to higher molecular weight PEG molecules due to the repeating characteristics of the backbone. The assay uses a monoclonal antibody to PEG pre-coated on a microtiter plate. The antibody binds in a competitive manner the PEG or PEGylated protein in the sample or the PEG covalently linked to biotin. After incubation at room temperature the excess reagents are washed away and a streptavidin HRP conjugate is added to bind to the biotinylated PEG bound to the antibody. After a second wash to remove excess HRP substrate is added. An HRP-catalyzed reaction gener-ates a blue color in the solution. The reaction is stopped with stop solution and the resulting yellow color is read at 450nm. The amount of signal is inversely proportional to the concentration of PEG.