全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...

猪繁殖与呼吸综合征病毒GP5蛋白中和性单克隆抗体的制备与免疫学特性测定

DOI: 10.7685/j.issn.1000-2030.2008.01.015, PP. 72-76

Keywords: 猪繁殖与呼吸综合征病毒(PRRSV),GP5蛋白,单克隆抗体,中和抗体

Full-Text   Cite this paper   Add to My Lib

Abstract:

利用重组真核质粒pcDNA3-GP5免疫BALB/c小鼠,取脾细胞和骨髓瘤细胞SP2/0融合,经间接ELISA筛选和3次有限稀释法克隆,得到3株能稳定分泌抗猪繁殖与呼吸综合征病毒(PRRSV)GP5蛋白的单克隆抗体(McAb)杂交瘤细胞株,分别命名为4C9、5D10、5F12,其中5D10的Ig亚类为IgM。ELISA检测杂交瘤细胞培养上清液抗体的效价为1∶64~1∶1024,腹水的效价为1∶3200~1∶10240,IFA和IPMA的检测结果均为阳性。Western-blot检测证明,这3株McAb均针对PRRSV的GP5蛋白。中和试验表明,5D10和5F12具有病毒中和活性,中和效价分别为1∶40和1∶80。5D10相对亲和力大于5F12,3株细胞连续培养20代后仍能稳定分泌抗体,表明抗GP5蛋白中和性McAb制备成功。

References

[1]  Pirzadeh B,Dea S.Monoclonal antibodies to the ORF5 product of porcine reproductive and respiratory syndrome virus define linear neutralizing determinants[J].Journal of General Virology.1997,78:1867-1873
[2]  Plagemann P G W,Rowland R R R,Faaberg K S.The primary neutralization epitope of porcine respiratory and reproductive syndrome virus strain VR-2332 is located in the middle of the GP5 ectodomain[J].Archives of Virology.2002,147:2327-2347
[3]  Zhang Y,Rameshwer D S,Prem S P.Monoclonal antibody against conformational dependent epitopes on porcine reproduction and respiratory syndrome[J].Veterinary Microbiology.1998,63:125-136
[4]  Yang L,Frey M L,Yoon K J.Categorization of North American procine reproductive and respiratory syndrome virus:epitopic profiles of the N,M,GP5 and GP3 proteins and susceptibility to neutralization[J].Archives of Virology.2000,145:1599-1619
[5]  白雪帆,杨为松,崔运昌.ELISA方法用于McAb亲和常数的测定[J].免疫学杂志.1992,8(2):126
[6]  Drew T W,Meulenberg J J M,Sands J J.Production characterization and reactivity of monoclonal antibodies to procine reproductive and respiratory syndrome virus[J].Journal of General Virology.1995,76(7):1361-1369
[7]  Cancel-Tirado S M,Evans R B,Yoon K J.Monoclonal antibody analysis of porcine reproductive and respiratory syndrome virus epitopes associated with antibody-dependent enhancement and neutralization of virus infection[J].Veterinary Immunology and Immunopathology.2004,102:249-262
[8]  蒋文明,姜平,李玉峰.猪繁殖与呼吸综合征病毒GP5蛋白N端信号肽对其DNA免疫应答的影响[J].病毒学报.2005,21(6):478-480
[9]  Yang L,Yoon K J,Li Y.Antigenic and genetic variations of the 15 kD nucleocapsid protein of porcine reproductive and respiratory syndrome virus isolates[J].Archives of Virology.1999,144:525-546
[10]  刘秀梵.单克隆抗体在农业上的应用[M].合肥:安徽科学技术出版社.1994:21-23;32-36
[11]  萨姆布鲁克?J,弗里奇 E F,曼尼阿蒂斯 T.分子克隆实验指南[M].北京:科学出版社.1992:27-62

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133