文献类型： 外文期刊

作者： Yang Yan-yan ¹ ; Zhu Li-qian ¹ ; Yang Ji-fei ¹ ; Teng Man ¹ ; Cai Shu-jun ¹ ; Wang Li ¹ ; Luo Jun ¹ ; Deng Rui-guang ¹ ; Zha ¹ ;

作者机构： 1.Henan Acad Agr Sci, Henan Prov Key Lab Anim Immunol, Zhengzhou 450002, Peoples R China

2.Yangzhou Univ, Coll Vet Med, Yangzhou 225009, Peoples R China

关键词： CEF;IBDV receptor;biotin-streptavidin

期刊名称：AGRICULTURAL SCIENCES IN CHINA （ 2013影响因子：0.82； 五年影响因子：0.997 ）

ISSN： 1671-2927

年卷期： 2010 年 9 卷 2 期

页码：

收录情况： SCI

摘要： This article aims to establish an efficient assay for screening monoclonal antibodies (McAbs) against the membrane proteins of chicken embryo fibroblast (CEF) for further studies of the cellular receptors of infectious bursal disease virus (IBDV). McAbs against the membrane proteins of CEF were prepared by cell fusion. The monolayer CEF pre-incubated with the CEF-specific McAbs for 2 h were infected with IBDV and incubated with F22-EA6-biotin postinfection. Then, the cells were reacted with streptavidin-horseradish peroxidase (HRP) and finally stained by 3-amino-9-ethylcarbazole (AEC). The inhibitive percentage of IBDV infection was calculated by counting the IBDV-infected cells to determine the inhibition efficiency of the CEF-specific McAbs. Compared with the control cells, the IBDV-infected cells pretreated with CEF-specific antibody significantly decreased; supernatant fluids of a total of 768 hybridomas were analyzed. The results of immunohistochemistry assays showed that six of them (1A5, 1H11, 2B12, 3G1, 4D10, and 4B8) have the abilities to block the infection of IBDV to CEF, among which 4B8 can perfectly block the infection. This novel method is a sensitive and specific assay for the screening of CEF membrane protein-specific McAbs, which can block the infection of IBDV to CEF, and these McAbs can be used for the further investigations of the cellular receptors of IBDV.