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

Analytical Biochemistry
Sang-Woo KimHyeon-Su Ro

Abstract

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 ...Continue Reading

References

Mar 21, 2003·Journal of Proteome Research·Cheng C WangJocelyn Burke
Jan 31, 2004·Analytical Biochemistry·Gabriela A CanzianiDavid G Myszka
Jun 19, 2004·Analytical Biochemistry·Jin-Mi JungBong Hyun Chung
Aug 3, 2005·Nature Reviews. Microbiology·Donald L Nuss
Dec 2, 2005·Nature·Jason PtacekMichael Snyder
Mar 3, 2006·Analytical Biochemistry·Pär SäfstenDavid G Myszka
May 29, 2007·Journal of Immunological Methods·Paul LeonardRichard O'Kennedy

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