本文選題：山羊　切入點：IFN-γ　出處：《西北農(nóng)林科技大學(xué)》2017年碩士論文

【摘要】：T細胞、NK細胞活化后,會產(chǎn)生干擾素-γ(IFN-γ),IFN-γ能夠增強巨噬細胞和中性粒細胞的殺傷活性,具有抗病毒感染、抗腫瘤等生物學(xué)活性。血液和炎癥部位IFN-γ含量的變化可作為評估機體細胞免疫功能的重要指標(biāo)。目前未見有山羊IFN-γ單抗的研究報道,也未見相關(guān)的試劑盒。為制備山羊IFN-γ單克隆抗體,本研究以重組的山羊IFN-γ融合蛋白為抗原免疫BALB/c小鼠,通過常規(guī)細胞融合等單克隆抗體制備技術(shù),篩選IFN-γ單抗雜交瘤,對IFN-γ單抗的特異性和亞型進行鑒定。研究結(jié)果如下:1.免疫的3只BALB/c小鼠血清抗體效價均達到1:105。2.對融合的雜交瘤細胞進行篩選,獲得了1株能夠分泌IFN-γ單抗的雜交瘤細胞株(命名為3C),雜交瘤細胞的培養(yǎng)上清中抗體效價為1:103,制備小鼠腹水后,測得抗體效價為1:106。3.IFN-γ單克隆抗體的類別為IgG1,κ型。4.在Western blot試驗中,IFN-γ單克隆抗體能與山羊IFN-γ重組蛋白發(fā)生特異性反應(yīng)。5.所獲得單抗能特異性識別PBMCs分泌的天然IFN-γ,能與患有臨床乳房炎的奶山羊乳汁中的IFN-γ發(fā)生特異性反應(yīng)。本研究獲得了1株能夠識別山羊IFN-γ的雜交瘤細胞,該雜交瘤細胞生長穩(wěn)定,抗體分泌量高。
[Abstract]:After T cell activation, IFN- 緯 -IFN- 緯 can enhance the killing activity of macrophages and neutrophils, and it has anti-virus infection. The changes of IFN- 緯 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- 緯 monoclonal antibody. In order to prepare goat IFN- 緯 monoclonal antibody, the recombinant goat IFN- 緯 fusion protein was used as antigen to immunize BALB/c mice. The specificity and subtype of IFN- 緯 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- 緯 monoclonal antibody was obtained. The antibody titer in the supernatant of hybridoma cell culture was 1: 103. The titer of monoclonal antibody against IFN- 緯 was 1: 106.3.IFN- 緯 was classified as IgG1, 魏 .4.In the Western blot test, the monoclonal antibody could react specifically with goat IFN- 緯 recombinant protein. The obtained monoclonal antibody could specifically recognize the natural IFN- 緯 secreted by PBMCs, and could specifically recognize the natural IFN- 緯 secreted by PBMCs. IFN- 緯 in milk of dairy goats with clinical mastitis was specifically reacted. In this study, a hybridoma cell line capable of recognizing goat IFN- 緯 was obtained. The hybridoma cells grow steadily and have high antibody secretion.
【學(xué)位授予單位】：西北農(nóng)林科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S852.4

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