Increased Sensitivity of an Enzyme Immunoassay for Human Interferon Alpha 1 Using a Mixture of Three Monoclonal Antibodies

摘要

A sensitive sandwich ELISA for the quantification of human interferon (IFN)-αl was designed. The assay employed IFN-αl- specific polyclonal antibody for coating and monoclonal antibodies (mAbs) to IFN-a 1 as a second antibody. A major increase in sensitivity of the assay could be achieved, when polyclonal antibody was combined with a mixture of three mAbs binding to different regions of IFN-al, compared to the combination of a polyclonal antibody with a single mAb. The sensitivity of the established ELISA was close to that of an antiviral IFN-bioassay. The ELISA did not cross-react with IFN-α2, β, τ or omega. The immunoassay allowed to estimate the content of IFN-αl in leukocyte iFN-α to about 25–50% or to 2–6% in Namalwa IFN-α, respectively.