[1]王 冰.赤羽病病毒N蛋白单克隆抗体的制备[J].中国预防兽医学报,2019,(03):309-312.[doi:0.3969/j.issn.1008-0589.201803018]
 WANG Bing.Preparation of the monoclonal antibodies against N protein of Akabane virus[J].Chinese journal of preventive veterinary medicine,2019,(03):309-312.[doi:0.3969/j.issn.1008-0589.201803018]
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赤羽病病毒N蛋白单克隆抗体的制备()
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2019年03
页码:
309-312
栏目:
出版日期:
2019-04-25

文章信息/Info

Title:
Preparation of the monoclonal antibodies against N protein of Akabane virus
文章编号:
1008-0589(2019)03-0309-04
作者:
 

王 冰

 (黑龙江农垦职业学院,黑龙江 哈尔滨 151025)
Author(s):
 

WANG Bing

 (Heilongjiang Agricultural Reclama tiun Vocational College, Harbin 151025, China)
关键词:
  关键词:赤羽病病毒N蛋白原核表达单克隆抗体
Keywords:
 

 Akabane virus N protein prokaryotic expression;  monoclonal antibody

分类号:
S852.65
DOI:
0.3969/j.issn.1008-0589.201803018
文献标志码:
A
摘要:
为制备赤羽病病毒(AKAV)N蛋白的单克隆抗体(MAb),本实验利用原核表达系统表达、纯化的重组N蛋白免疫BALB/c小鼠,三免后取小鼠脾淋巴细胞与SP2/0细胞融合,并以纯化的重组N蛋白为包被抗原,通过间接ELISA筛选出1株稳定分泌抗AKAV N蛋白的MAb杂交瘤细胞株IIG5,检测结果显示IIG5为IgG1/κ链。间接免疫荧光结果显示,IIG5与AKAV感染细胞呈阳性反应,与茨城病病毒、中山病病毒以及蓝舌病病毒1型均呈阴性反应,特异性较好。Western blot结果显示,IIG5能够识别重组的以及AKAV感染细胞中的N蛋白。本研究结果为AKAV检测方法的建立及相关研究奠定基础。
 
Abstract:
For the preparation of monoclonal antibodies (MAbs) against N protein of Akabane virus (AKAV), SP2/0 myeloma cells were fused with spleen cells from BALB/c mice, which were immunized with purified recombinant N protein of AKAV expressed by E.coli expression system. One hybridoma cell lines stably secreting MAbs against N protein of AKAV, namly IIG5, were identified by indirect ELISA coated with the purified recombinant N protein. Indirect immunofluorescence assay indicated that IIG5 was able to react positively with AKAV infected cells, and no cross reactions with Ibaraki virus, Chuzan disease virus and serum type 1 Bluetongue virus were observed. Western blot identification showed IIG5 also reacted with the recombinant N protein and N protein from AKAV infected cells. The study could help to develop AKAV type-specific detection method and other associated research.

参考文献/References:

[1] Sevik M. Molecular detection and genetic analysis of Akabane virus genogroup Ib in small ruminants in Turkey [J]. Arch Virol, 2017, 6: doi: 10.1007/s00705-017-3398-x.
[2]Tohru Y, Tomoko K, Yoko H, et al. Oral susceptibility of Japanese Culicoides (Diptera: Ceratopogonidae) species to Akabane virus [J]. J Med Entomol, 2019, 56(2): 533-539.
[3]Lim S I, Kweon C H, Tark D S, et al. Sero-survey on Aino, Akabane, Chuzan, bovine ephemeral  fever and Japanese encephalitis virus of cattle and swine in Korea [J]. J Vet Sci, 2007, 8(1): 45-49.
[4]Feng Yun, Zhang Yu-zhen, Yang Wei-hong, et al. Characterization and analyses of the full-length genome of a strain of the Akabane virus isolated from Mosquitoes in Yunnan province, China [J]. Bing Du Xue Bao, 2016, 32(2): 161-169.
[5]Kirkland P D. Akabane virus infection [J]. Rev Sci Tech, 2015, 34(2): 403-410.
[6]Zhang Yong-ning, Wu Shao-qiang, Wang Jian-chang, et al. Expression and purification of the nucleocapsid protein of Schmallenberg virus, and preparation and characterization of a monoclonal antibody against this protein [J]. Protein Expr Purif, 2013, 92(1): 1-8.
[7]Zhang Gai-ping, Li Ning, Chen Yu-mei, et al. Identification of the B-cell epitopes on N protein of type 2 porcine reproductive and respiratory syndrome virus, using monoclonal antibodies[J]. Int J Biol Macromol, 2019, 130(18): 300-306.
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                                        (本文编辑:李 娜)

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更新日期/Last Update: 2019-04-26