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Published on: May 6, 2015
[Preparation, characterization and application of anti-human adenovirus monoclonal antibody with neutralization
Si-Xiu Li1, Guo-Cheng Zhang, Dong-Liang Xu
1Department of Pediatrics, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China.
Aim:
To prepare, identify and apply anti-human adenoviru(HAdv)neutralization monoclonal antibody(mAb).
Methods:
BALB/c mice were immunized with live human adenovirus type3(HAdv-3) strain intranarially. Sp2/0 cells were fused with the spleen cells harvested from BALB/c mice. The chromosomal amounts of the hybridoma cells were analyzed by colchicine. A commercially available mouse mAb isotyping kit was used to identify the isotype of this mAb. Clones secreting specific monoclonal antibody were screened by indirect enzyme linked immunosorbent assay (ELISA), Western blot and indirect immunofluorescent assay. Infected animal model was established, and the protective effect of mAb was studied.
Results:
The fusion rate was 86%, and the positive rate was 51.4%. One of the hybridoma cell was identified(1A4), the chromosomal amounts of was 98, the subtype mAb type belonged to IgG2a/kappa, and the titer of the mAb secreted by the strain in ascite reached more than 10(-5);. The specificity of the mAb was proved by ELISA , Western blot and indirect immunofluorescent assay. This mAb had protective effect on animal infected by HAdv-3.
Conclusion:
The anti-adenovirus mAb which have neutralization activities was successfully prepared. The mAb recognized the hexon subunit and had protective effect on animal infected by HAdv-3.
