Abstract
Aim:
To transform the human anti-rabies virus glycoprotein (anti-RABVG) single-chain variable fragment (scFv) into a Fab fragment and to analyze its immunological activity.
Methods:
The Fab gene was amplified using overlap PCR and inserted into the vector pComb3XSS. The recombinant vector was then transformed into E coli Top10F' for expression and purification. The purified Fab was characterized using SDS-PAGE, Western blotting, indirect ELISA, competitive ELISA, and the fluorescent antibody virus neutralization test (FAVN), respectively, and examined in a Kunming mouse challenge model in vivo.
Results:
A recombinant vector was constructed. The Fab was expressed in soluble form in E coli Top10F'. Specific binding of the Fab to rabies virus was confirmed by indirect ELISA and immunoprecipitation (IP). The neutralizing antibody titer of Fab was 10.26 IU/mL. The mouse group treated with both vaccine and human rabies immunoglobulin (HRIG)/Fab091 (32 IU/kg) showed protection against rabies, compared with the control group (P<0.05, Logrank test).
Conclusion:
The antibody fragment Fab was shown to be a neutralizing antibody against RABVG. It can be used together with other monoclonal antibodies for post-exposure prophylaxis of rabies virus in future studies.
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Acknowledgements
This investigation was supported by the National High Technology Research and Development Program, China (No 2007AA02Z418). We would like to thank Chun-yan GU for valuable technical assistance and Johnson HUANG for revising the manuscript.
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Li, C., Zhang, F., Lin, H. et al. Generation and characterization of the human neutralizing antibody fragment Fab091 against rabies virus. Acta Pharmacol Sin 32, 329–337 (2011). https://doi.org/10.1038/aps.2010.209
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DOI: https://doi.org/10.1038/aps.2010.209
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