本文選題：犬瘟熱病毒　切入點(diǎn)：單克隆抗體　出處：《中國(guó)預(yù)防獸醫(yī)學(xué)報(bào)》2017年07期 　論文類(lèi)型：期刊論文

【摘要】：為制備犬瘟熱病毒(CDV)單克隆抗體(MAb),本研究用腺病毒表達(dá)的CDV截短N(yùn)蛋白(aa401~aa523)免疫BALB/c小鼠,取免疫小鼠脾細(xì)胞與SP2/0骨髓瘤細(xì)胞進(jìn)行融合并篩選到2株能夠穩(wěn)定分泌抗CDV N蛋白的MAb,命名為N1-C8和N1-C41。經(jīng)間接ELISA測(cè)定N1-C8和N1-C41培養(yǎng)上清液及小鼠腹水效價(jià)分別為1∶1 024、1∶106和1∶512、1∶105。通過(guò)肽掃描的方法篩選出N1-C8的抗原表位為線性表位~(440)ENQGGDKYPIHFNDE454,但并未能夠篩選出N1-C41的抗原表位,推測(cè)可能是因?yàn)槠淇乖砦粸榭臻g構(gòu)象型。Western blot結(jié)果顯示N1-C8和N1-C41兩株MAb在識(shí)別CDV的不同毒力病毒株上有差異,可能與CDV強(qiáng)弱病毒株在N蛋白的差異變化有關(guān)。本研究?jī)芍闙Ab的制備為CDV不同毒力株的鑒別診斷提供了可能。
[Abstract]:In order to prepare a monoclonal antibody against canine distemper virus (CDV), BALB/c mice were immunized with CDV truncated N protein (aa401a 523) expressed by adenovirus. The spleen cells of immunized mice were fused with SP2/0 myeloma cells, and two strains of MAbs which could stably secrete anti CDV N protein were selected and named N1-C8 and N1-C41.The titers of supernatants and ascites of N1-C8 and N1-C41 cultured in mice were determined by indirect ELISA, and the titers of supernatants and ascites of mice were 1: 10241: 106, respectively. The antigen epitopes of N1-C8 were obtained by peptide scanning. The epitope of N1-C8 was a linear epitope, ENQGDDKYPIHFNDE4544.However, the epitopes of N1-C41 could not be screened out. It is suggested that the antigenic epitopes of N1-C8 and N1-C41 have different virulence in different virulent strains of CDV. The results of Western blot indicate that the antigenic epitopes of N1-C8 and N1-C41 are different in different virulence virus strains. It may be related to the variation of N protein in CDV strain. The preparation of two strains of MAb provides the possibility for differential diagnosis of different virulence strains of CDV.
【作者單位】： 中國(guó)農(nóng)業(yè)科學(xué)院特產(chǎn)研究所特種經(jīng)濟(jì)動(dòng)物分子生物學(xué)國(guó)家重點(diǎn)實(shí)驗(yàn)室;
【基金】：吉林省自然科學(xué)基金(20150101104JC)
【分類(lèi)號(hào)】：S852.65

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