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- Title
Preparation and Evaluation of Human-Murine Chimeric Antibody against Protective Antigen of Bacillus anthracis.
- Authors
Lina Hao; Feng Zheng; Siping Xiong; Dan Hu; Heng Lv; Qi Tang; Jin Yang; Zhenqing Feng; Changjun Wang; Jin Zhu
- Abstract
The aim of this research is to develop a human/murine chimeric Fab antibody which neutralizes the anthrax toxin, protective antigen (PA). The chimeric Fab was constructed using variable regions of murine anti-PA monoclonal antibody in combination with constant regions of human IgG. The chimeric PA6-Fab was expressed in E. coli. BL21 and evaluated by ELISA and co-immunoprecipitation- mass spectra. The potency of PA6-Fab to neutralize LeTx was examined in J774A.1 cell viability in vitro and in Fisher 344 rats in vivo. The PA6-Fab did not have domain similarity corresponding to the current anti PA mAbs, but specifically bound to anthrax PA at an affinity of 1.76 nM, and was able to neutralize LeTx in vitro and protected 56.9% cells at 20 pg/mL against anthrax LeTx. One hundred pg PA6-Fab could neutralize 300 pg LeTx in vivo. The PA6-Fab has potential as a therapeutic mAb for treatment of anthrax.
- Subjects
IMMUNOGLOBULIN synthesis; ANTHRAX toxin; BACILLUS anthracis; ANTIGENS; CHIMERIC proteins; CELL survival; IMMUNOPRECIPITATION
- Publication
International Journal of Molecular Sciences, 2014, Vol 15, Issue 10, p18496
- ISSN
1661-6596
- Publication type
Article
- DOI
10.3390/ijms151018496