[1]高 冉,刘建波,陈建飞,等.抗猪SIgA SC片段单克隆抗体的制备及鉴定[J].中国预防兽医学报,2018,(05):455-458.[doi:0.3969/j.issn.1008-0589.201707014]
 GAO Ran,LIU Jian-bo,CHEN Jian-fei,et al. Preparation and identification of monoclonal antibodies against porcine SIgA SC fragment[J].Chinese journal of preventive veterinary medicine,2018,(05):455-458.[doi:0.3969/j.issn.1008-0589.201707014]
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抗猪SIgA SC片段单克隆抗体的制备及鉴定()
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2018年05
页码:
455-458
栏目:
研究简报
出版日期:
2018-05-15

文章信息/Info

Title:
 

Preparation and identification of monoclonal antibodies against porcine SIgA SC fragment

文章编号:
1008-0589(2018)5-0455-04
作者:
 

高 冉12刘建波2陈建飞2时洪艳2张 鑫2石 达2曹丽艳2董素凤3冯 力2*

 (1. 黑龙江八一农垦大学 动物科技学院,黑龙江 大庆 163319;2. 中国农业科学院哈尔滨兽医研究所 兽医生物技术国家重点实验室/猪消化道团队,黑龙江 哈尔滨 150069;3. 辽宁省重大动物疫病应急中心,辽宁 沈阳 110161)
Author(s):
GAO Ran12 LIU Jian-bo2 CHEN Jian-fei2 SHI Hong-yan2 ZHANG Xin2 SHI Da2 CAO Li-yan2DONG Su-feng3 FENG Li2*
  (1. College of Animal Science and Technology, Heilongjiang Bayi Agricultural University, Daqing 163319, China; 2. Pig Gastrointestinal Tract Team, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, China; 3. Major Animal Epidemic Emergency Center in Liaoning Province, Shenyang 110161, China)
关键词:
SIgA单克隆抗体SC片段
Keywords:
secretory IgA  monoclonal antibody  secretory component
分类号:
S852.4
DOI:
0.3969/j.issn.1008-0589.201707014
文献标志码:
A
摘要:
为获得抗猪分泌型IgA (SIgA)的单克隆抗体(MAb),本研究以纯化的SIgA蛋白为抗原,免疫BALB/c小鼠,取其脾细胞与SP2/0细胞融合,通过间接ELISA方法筛选出1株稳定分泌抗SIgA SC片段的杂交瘤细胞株,命名为2F9。MAb亚类鉴定结果显示其亚类为IgG1/κ链。杂交瘤细胞培养上清液和腹水的效价分别为1∶200和1∶16 000。Western blot鉴定显示该MAb能够识别天然SIgA蛋白;IFA显示该MAb可以识别真核细胞表达的SIgA SC蛋白。抗猪SIgA SC片段的MAb制备,为建立一系列猪消化道病原的特异性SIgA诊断方法及研究黏膜免疫提供了重要工具,具有广阔的应用前景。
  
Abstract:
To prepare the MAb against porcine SIgA, the purified SIgA was used as antigen to immunize BALB/c mice. Then the spleen cells of the immunized mice were fused with SP2/0 cells. One hybidoma cell line stably secreting MAb against SIgA SC protein was identified by indirect ELISA, named 2F9. The MAb belonged to IgG1 subtype with κ chain. The titers in cell culture medium of hybridoma cell and ascites were 1∶200 and 1∶16 000, respectively. Western blot assay showed that native SIgA could be identified by 2F9. IFA assay showed that eukaryotic expressed SC protein could be identified by 2F9. The preparation of the MAb would provide an important tool for establishing a series of diagnosis methods to detect porcine digestive tract pathogens based on specific SIgA and for further study on mucosal immunity. The preparation of SC MAb has broad application prospects.

参考文献/References:

[1]Chairatana P,  Nolan E M. Defensins, Lectins, Mucins and secretory immunoglobulin A: microbe-binding biomolecules that contribute to mucosal immunity in the human Gut [J]. Biochem Mol Biol, 2017, 52(1): 45-56.
[2]Brandtzaeg P. Secretory IgA: designed for anti-microbial defense [J]. Front Immunol, 2013, (4): 1-17.
[3] Song Wei-xiang, Feng Zhi-xin, Bai Yun, et al. Preparation of the porcine secretory component and a monoclonal antibody against this protein [J]. Protein Expr Purif, 2015, (113): 51-55.
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                               (本文编辑:李 娜)

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更新日期/Last Update: 2018-05-16