抗山羊IFN-γ单抗的制备及鉴定

发布时间：2018-03-23 16:20

  本文选题：山羊　切入点：IFN-γ　出处：《西北农林科技大学》2017年硕士论文

【摘要】：T细胞、NK细胞活化后,会产生干扰素-γ(IFN-γ),IFN-γ能够增强巨噬细胞和中性粒细胞的杀伤活性,具有抗病毒感染、抗肿瘤等生物学活性。血液和炎症部位IFN-γ含量的变化可作为评估机体细胞免疫功能的重要指标。目前未见有山羊IFN-γ单抗的研究报道,也未见相关的试剂盒。为制备山羊IFN-γ单克隆抗体,本研究以重组的山羊IFN-γ融合蛋白为抗原免疫BALB/c小鼠,通过常规细胞融合等单克隆抗体制备技术,筛选IFN-γ单抗杂交瘤,对IFN-γ单抗的特异性和亚型进行鉴定。研究结果如下:1.免疫的3只BALB/c小鼠血清抗体效价均达到1:105。2.对融合的杂交瘤细胞进行筛选,获得了1株能够分泌IFN-γ单抗的杂交瘤细胞株(命名为3C),杂交瘤细胞的培养上清中抗体效价为1:103,制备小鼠腹水后,测得抗体效价为1:106。3.IFN-γ单克隆抗体的类别为IgG1,κ型。4.在Western blot试验中,IFN-γ单克隆抗体能与山羊IFN-γ重组蛋白发生特异性反应。5.所获得单抗能特异性识别PBMCs分泌的天然IFN-γ,能与患有临床乳房炎的奶山羊乳汁中的IFN-γ发生特异性反应。本研究获得了1株能够识别山羊IFN-γ的杂交瘤细胞,该杂交瘤细胞生长稳定,抗体分泌量高。
[Abstract]:After T cell activation, IFN- 纬 -IFN- 纬 can enhance the killing activity of macrophages and neutrophils, and it has anti-virus infection. The changes of IFN- 纬 content in blood and inflammatory site can be used as an important index to evaluate the cellular immune function of the body. There are no studies on goat IFN- 纬 monoclonal antibody. In order to prepare goat IFN- 纬 monoclonal antibody, the recombinant goat IFN- 纬 fusion protein was used as antigen to immunize BALB/c mice. The specificity and subtype of IFN- 纬 monoclonal antibody were identified. The results were as follows: 1. The titers of serum antibodies of three BALB/c mice immunized were 1: 105.2. The fusion hybridoma cells were screened. A hybridoma cell line that secreted IFN- 纬 monoclonal antibody was obtained. The antibody titer in the supernatant of hybridoma cell culture was 1: 103. The titer of monoclonal antibody against IFN- 纬 was 1: 106.3.IFN- 纬 was classified as IgG1, 魏 .4.In the Western blot test, the monoclonal antibody could react specifically with goat IFN- 纬 recombinant protein. The obtained monoclonal antibody could specifically recognize the natural IFN- 纬 secreted by PBMCs, and could specifically recognize the natural IFN- 纬 secreted by PBMCs. IFN- 纬 in milk of dairy goats with clinical mastitis was specifically reacted. In this study, a hybridoma cell line capable of recognizing goat IFN- 纬 was obtained. The hybridoma cells grow steadily and have high antibody secretion.
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S852.4

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