Identification and primary application of hybridomas cell secreting monoclonal antibodies against mink (Neovison vison) interferon-gamma

文献类型： 外文期刊

第一作者： Zhang, Hailing

作者： Zhang, Hailing;Fan, Sining;Zhang, Lei;Hu, Bo;Bai, Xue;Lu, Rongguang;Zhao, Jianjun;Lian, Shizhen;Gao, Bing;Yan, Xijun;Zhu, Yanzhu;Zhang, Hailing;Zhang, Shasha;Lu, Shiying;Zhang, Dongliang

作者机构：

关键词： Mink; Interferon-gamma; Monoclonal antibody; Hybridoma clones; Lymphocyte; ELISA method

期刊名称：CYTOKINE （ 影响因子：3.926； 五年影响因子：4.033 ）

ISSN： 1043-4666

年卷期： 2022 年 150 卷

页码：

收录情况： SCI

摘要： Due to their susceptibility to several human viruses, the mink has been proposed as potential animal models for the study of human viral infections. However, there are no specific monoclonal antibody (mAbs) currently available for the detection of mink-specific interferon-gamma (miIFN-gamma). The BALB/c mice were immunized intraperitoneally with purified recombinant miIFN-gamma protein. The splenocytes were obtained and fused with murine myeloma cells. Five of 24 hybridoma clones were obtained to produce mAbs steadily with the strongest affinity to recombinant miIFN-gamma protein. The isotype of the 31A, 31B and 31G were lgG 2b. The isotype of 44 and 46 were lgG 2a and 1. All five mAbs were. light chains. Western blotting and indirect ELISA method showed that 5 mAbs were positive to miIFN-gamma. Immunofluorescence showed that 2 mAbs (44 and 46) had a positive reaction to miIFN-gamma. The hybridoma clone 46 had the highest sensitivity for the detection of miIFN-gamma. Most importantly, our primary sandwich ELISA system (mAbs 46 and polyclonal antiserum) detected endogenous IFN-gamma in mink lymphocytes infected with canine distemper virus (CDV). We have thus developed a novel mAbs could recognize miIFN-gamma, and have demonstrated the first ELISA-based measurement of IFN-gamma in lymphocyte of the mink.

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