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


CONCLUSIONS
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.
Zhu
Yin-hua
YH
Department of Forensic Biology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
Liang
Wei-bo
WB
Lü
Me-li
ML
Xu
Yong-chun
YC
Jia
Jing
J
Zhang
Lin
L
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000096:biosynthesis*; D005260:Female; D005982:Glutathione Transferase / Q000096:biosynthesis / Q000235:genetics*; D004854:Herpesvirus 4, Human / Q000235:genetics; D017874:Immediate-Early Proteins / Q000096:biosynthesis / Q000235:genetics*; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D015534:Trans-Activators / Q000096:biosynthesis / Q000235:genetics*; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics*
D000911:Antibodies, Monoclonal; C068382:BRLF1 protein, Human herpesvirus 4; D017874:Immediate-Early Proteins; D011993:Recombinant Fusion Proteins; D015534:Trans-Activators; D014764:Viral Proteins; D005982:Glutathione Transferase