An application of protein microarray in the screening of monoclonal antibodies against the oyster mushroom spherical virus

作     者：Kim, Sang-Woo Kim, Min-Gon Jung, Hyo-Am Lee, Kyung-Hee Lee, Hyun-Sook Ro, Hyeon-Su 

作者机构：Gyeongsang Natl Univ Dept Microbiol Chinju 660701 South Korea Gyeongsang Natl Univ Res Inst Life Sci Chinju 660701 South Korea Korea Res Inst Biosci & Biotechnol BioNanotechnol Res Ctr Taejon 305600 South Korea Gyeongsang Natl Univ Environm Biotechnol Natl Core Res Ctr Chinju 660701 South Korea 

出 版 物：《ANALYTICAL BIOCHEMISTRY》 (分析生物化学)

年 卷 期：2008年第374卷第2期

页      面：313-317页

核心收录：

学科分类：0710[理学-生物学] 071010[理学-生物化学与分子生物学] 07[理学] 0703[理学-化学] 

基　　金：Environmental Biotechnology National Core Research Center Korea Industrial Technology Foundation, (PS-06-1-008) Korea Science and Engineering Foundation, KOSEF Ministry of Science and Technology, Taiwan, MOST 

主　　题：protein microarray monoclonal antibody mycovirus screening fungi 

摘      要：The oyster mushroom spherical virus (OMSV) is a causative agent of dieback disease in the oyster mushroom, Pleurotus ostreatus. Outbreaks of this virus occasionally result in serious disease that is associated with hefty economic losses. Thus, the detection and removal of OMSV-infected spawn is considered to be a crucial step for the stable production of P. ostreatus. For the detection of OMSV, we attempted to generate monoclonal antibodies (mAbs) against an RNA polymerase domain (RPD) of an OMSV protein. In an effort to simplify the laborious multistep mAb screening process, we developed a protein microarray on a slide glass that is chemically modified with the RPD protein. The culture supernatants of 87 hybridoma cells, which were prepared from the fusion of RPD-immunized mouse spleen cells with myeloma cells, were spotted onto the RPD-coated microarray. The binding of mAb to RPD was detected via Alexa 488 dye-labeled anti-mouse immunoglobulin G (IgG) as a secondary antibody. Of 87 samples, 13 evidenced a significant level of fluorescence signal intensity. Subsequent immunoblot analysis revealed that the specificity of each mAb against RPD coincided with the corresponding fluorescence signal intensity, thereby indicating the effectiveness of the protein microarray in mAb screening. (C) 2007 Elsevier Inc. All rights reserved.