Background: This ELISA detects antibodies against PEG-asparaginase, including responses directed toward the PEG moiety or the enzyme itself. Immunogenicity is a major concern with asparaginase therapies due to potential silent inactivation and hypersensitivity reactions. The assay supports clinical immunogenicity monitoring in oncology settings.
Principle Of Assay: This ELISA is a sandwich immunoassay. Antibodies are coated on 96 well plates. The antigen protein present in sample and standard respectively bind to the coated wells. The wells are washed and an antibody:HRP Conjugate is added which binds to the bound complex in the well. Washing is performed to remove any unbound material. TMB substrate is added and the enzyme reaction is stopped by dispensing of stop solution into the wells. The optical density (OD) of the solution at 450 nm is directly proportional to the amount of antigen protein present in the standard or samples.