[Preparation and identification of monoclonal antibody against human GST-Rta150 fusion protein from Epstein-Barr virus].

ABSTRACT

OBJECTIVE: To prepare a monoclonal antibody (mAb) against GST-Rta150 fusion protein from EB virus and identify its characteristics. METHODS: BALB/c mouse was rendered immune by GST-Rta150 fusion protein; after that, the spleen cells of immunized mouse were taken to fuse with SP2/0 myeloma cells by a routine method. Then screening for positive hybridoma cells and subclone. Following that, the hybridoma cells were injected into the peritoneal cavity of BALB/c mouse. The ascites was collected and purified. The prepared mAb was identified by double immunodiffusion method for its subclass; in addition, its titer and specificity were identified by ELISA and Western-blot, respectively. RESULTS: One hybridoma cell line 1A9 which stably secreted mAb against GST-Rta150 fusion protein was obtained. Its Ig subclass belonged to IgG1, the titer degree of the purified mAb being 10(-6). Western-blot analysis showed the mAb could combine with GST-Rta150 fusion protein specially. The titer degree of this hybridoma cell line remained unchanged after frozen in liquid nitrogen (LN) for three months. CONCLUSION: The mAb obtained by this method has powerful specificity and high stability. This work could serve as a foundation on which to study the Rta protein from EB virus and the diagnosis and treatment of EB virus correlated diseases.