Application of four anti-human interferon-alpha monoclonal antibodies for immunoassay and comparative analysis of natural interferon-alpha mixtures.
Four different mouse monoclonal antibodies to human interferon-alpha (IFN-alpha) were evaluated for application in quantitative and comparative analysis of natural IFN-alpha mixtures. Binding to IFN-alpha subtypes in solution revealed individual reactivity patterns. These patterns changed if the IFN-alpha molecules were immobilized either passively to a surface or bound by another antibody. Also, substitution of a single amino acid in IFN-alpha 2 affected the binding, apparently by altering the conformation. Isoelectric focusing of three natural IFN-alpha preparations from different sources, followed by immunoblotting, resulted in individual patterns with each of the four mAbs and also demonstrated variation in the composition of the IFN-alpha preparations. None of the mAbs was subtype specific, but by combining the different mAbs, and also applying polyclonal anti-human IFN-alpha antibodies, it was possible to design sensitive sandwich ELISAs with broad or more limited IFN-alpha subtype specificity.
Andersson
G
G
Research and Development Immunobiology, Kabi Biopharma, Stockholm, Sweden.
Lundgren
E
E
Ekre
H P
HP
D000327:Adsorption; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D001681:Biological Assay; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D015151:Immunoblotting; D007370:Interferon Type I / Q000032:analysis* / Q000276:immunology; D007457:Iodine Radioisotopes; D007525:Isoelectric Focusing; D011994:Recombinant Proteins; D012996:Solutions
D000911:Antibodies, Monoclonal; D007370:Interferon Type I; D007457:Iodine Radioisotopes; D011994:Recombinant Proteins; D012996:Solutions