Sattayasai N, Marzuki S, McMullen G L, Geysen H M, Mason T J, Hibbs A R, Overall M, Hertzog P J, Linnane A W
Department of Biochemistry, Monash University, Clayton, Victoria, Australia.
Mol Immunol. 1991 Sep;28(9):975-83. doi: 10.1016/0161-5890(91)90183-k.
A strategy for the production of human interferon-alpha (IFN-alpha) 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-alpha 1, 50-57 and 103-116 of IFN-alpha 2, and 37-50 of IFN-alpha 4. The antipeptide antibodies [anti-IFN alpha 1(99-111), anti-IFN alpha 2(50-57C), anti-IFN alpha 2(103-116) and anti-IFN alpha 4(C37-50)] were tested by ELISA and Western blotting for their reactivity with immunoaffinity-purified recombinant human IFN-alpha 1, -alpha 2b and -alpha 4a. The anti-IFN alpha 1(99-111) and anti-IFN alpha 2(50-57C) reacted with their corresponding IFN-alpha and did not crossreact with the other IFN subtypes. The anti-IFN alpha 2(103-116) reacted with IFN-alpha 2b and also crossreacted slightly with the other subtypes. The anti-IFN alpha 4(C37-50) reacted well with IFN-alpha 4a, crossreacted with significantly lower affinity with IFN-alpha 1 and did not bind IFN-alpha 2b. Residues 104-107 and 108-111 are the major components of the epitopes recognized by anti-IFN alpha 1(99-111) and anti-IFN alpha 2(103-116), respectively, as determined by ELISA against overlapping octapeptides.
一种基于用与蛋白质载体偶联的短独特合成肽免疫兔子来生产人α干扰素（IFN-α）亚型特异性抗体的策略已得到验证。这些肽对应于IFN-α1的第99 - 111位氨基酸残基、IFN-α2的第50 - 57位和第103 - 116位氨基酸残基以及IFN-α4的第37 - 50位氨基酸残基。通过酶联免疫吸附测定（ELISA）和蛋白质印迹法检测抗肽抗体[抗IFNα1(99 - 111)、抗IFNα2(50 - 57C)、抗IFNα2(103 - 116)和抗IFNα4(C37 - 50)]与免疫亲和纯化的重组人IFN-α1、-α2b和-α4a的反应性。抗IFNα1(99 - 111)和抗IFNα2(50 - 57C)与其相应的IFN-α反应，且不与其他IFN亚型发生交叉反应。抗IFNα2(103 - 116)与IFN-α2b反应，也与其他亚型有轻微交叉反应。抗IFNα4(C37 - 50)与IFN-α4a反应良好，与IFN-α1的交叉反应亲和力显著较低，且不结合IFN-α2b。通过针对重叠八肽的ELISA测定，第104 - 107位和第108 - 111位残基分别是抗IFNα1(99 - 111)和抗IFNα2(103 - 116)识别的表位的主要组成部分。
