Increased sensitivity of an enzyme immunoassay for human interferon alpha 1 using a mixture of three monoclonal antibodies.
A sensitive sandwich ELISA for the quantification of human interferon (IFN)-alpha 1 was designed. The assay employed IFN-alpha 1- specific polyclonal antibody for coating and monoclonal antibodies (mAbs) to IFN-alpha 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-alpha 1, 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-alpha 2, beta, tau or omega. The immunoassay allowed to estimate the content of IFN-alpha 1 in leukocyte IFN-alpha to about 25-50% or to 2-6% in Namalwa IFN-alpha, respectively.
Karayianni-Vasconcelos
G
G
Institute of Virology, Slovak Academy of Sciences, Bratislava.
Mucha
V
V
Kontsek
P
P
D000911:Antibodies, Monoclonal / Q000276:immunology*; D001681:Biological Assay; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D016898:Interferon-alpha / Q000032:analysis / Q000276:immunology*; D009500:Neutralization Tests; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D016898:Interferon-alpha