Subtype-specificity of antipeptide antibodies raised against unique sequences of human interferons-α

Nison Sattayasai, Sangkot Marzuki, Gabrielle L. McMullen, H. Mario Geysen, Tom J. Mason, Alan R. Hibbs, Maree Overall, Paul J. Hertzog, Anthony W. Linnane

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A strategy for the production of human interferon-α (IFN-α) subtype-specific antibodies, based on immunizing rabbits with short unique synthetic peptides coupled to protein carriers, has been validated. These peptides correspond to amino acid residues 99-111 of IFN-α 1, 50-57 and 103-116 of IFN-α2, and 37-50 of IFN-α4. The antipeptide antibodies anti-IFNα 1(99-111), anti-IFNα2(50-57C), anti-IFNα2(103-116and anti-IFNα4(C37-50) were tested by ELISA and Western blotting for their reactivity with immunoaffinitypurified recombinant human IFN-α 1, -α2b and -α4a. The anti-IFNα 1(99-111) and anti-IFNα 2(50-57C) reacted with their corresponding IFN-α and did not crossreact with the other IFN subtypes. The anti-IFNα 2(103-116) reacted with IFN-α 2b and also crossreacted slightly with the other subtypes. The anti-IFNα 4(C37-50) reacted well with IFN-α 4a, crossreacted with significantly lower affinity with IFN-α 1 and did not bind IFN-α2b. Residues 104-107 and 108-111 are the major components of the epitopes recognized by anti-IFNα 1(99-111) and anti-IFNα2(103-116), respectively, as determined by ELISA against overlapping octapeptides.

Original languageEnglish
Pages (from-to)975-983
Number of pages9
JournalMolecular Immunology
Issue number9
Publication statusPublished - 1 Jan 1991

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