Comparison of antigenic properties of three interferon (IFN)-alpha 2 subvariants and establishement of a quantitative IFN-alpha 2 ELISA.
Three subvariants of human IFN-alpha 2 (2a, 2b, 2c) were found antigenically highly homologous, using a panel of specific monoclonal antibodies (MoAbs) directed to seven epitopes. Only in the region 30-41 IFN-alpha 2c showed some difference from the corresponding structures of subvariants 2a and 2b. An universal sandwich ELISA for the quantification of all subvariants of human IFN-alpha 2 was designed. A major increase in sensitivity of the immunoassay could be achieved, when polyclonal antibody to IFN-alpha 2 was combined with a mixture of three MoAbs to distinct sites of IFN-alpha 2, compared to the combination of a polyclonal antibody with a single MoAb. The sensitivity of the established ELISA ranged between 1-10 units/ml of IFN-alpha 2 and no cross-reactivity with IFN-alpha 1, -beta, or -omega 1 could be observed. We estimated the content of IFN-alpha 2 to about 56% in leukocyte IFN-alpha or to about 72% in Namalwa IFN-alpha.
Karayianni-Vasconcelos
G
G
Institute of Virology, Slovak Academy of Sciences, Bratislava.
Fuchsberger
N
N
Skamlová
Z
Z
Kontsek
P
P
D000818:Animals; D000906:Antibodies; D000911:Antibodies, Monoclonal; D000940:Antigenic Variation / Q000276:immunology; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D016898:Interferon-alpha / Q000032:analysis / Q000145:classification / Q000276:immunology*; D012680:Sensitivity and Specificity; D012756:Sheep
D000906:Antibodies; D000911:Antibodies, Monoclonal; D016898:Interferon-alpha