fj.12-219964.pdf (320.64 kB)
Production, characterization, and antigen specificity of recombinant 62-71-3, a candidate monoclonal antibody for rabies prophylaxis in humans
journal contribution
posted on 2023-06-09, 14:32 authored by Leonard Both, Craig van Dolleweerd, Edward WrightEdward Wright, Ashley C Banyard, Bianca Bulmer-Thomas, David Selden, Friedrich Altmann, Anthony R Fooks, Julian K C MaRabies kills many people throughout the developing world every year. The murine monoclonal antibody (mAb) 62-71-3 was recently identified for its potential application in rabies postexposure prophylaxis (PEP). The purpose here was to establish a plant-based production system for a chimeric mouse-human version of mAb 62-71-3, to characterize the recombinant antibody and investigate at a molecular level its interaction with rabies virus glycoprotein. Chimeric 62-71-3 was successfully expressed in Nicotiana benthamiana. Glycosylation was analyzed by mass spectroscopy; functionality was confirmed by antigen ELISA, as well as rabies and pseudotype virus neutralization. Epitope characterization was performed using pseudotype virus expressing mutagenized rabies glycoproteins. Purified mAb demonstrated potent viral neutralization at 500 IU/mg. A critical role for antigenic site I of the glycoprotein, as well as for two specific amino acid residues (K226 and G229) within site I, was identified with regard to mAb 62-71-3 neutralization. Pseudotype viruses expressing glycoprotein from lyssaviruses known not to be neutralized by this antibody were the controls. The results provide the molecular rationale for developing 62-71-3 mAb for rabies PEP; they also establish the basis for developing an inexpensive plant-based antibody product to benefit low-income families in developing countries.
History
Publication status
- Published
File Version
- Published version
Journal
The FASEB JournalISSN
0892-6638Publisher
Federation of American Society of Experimental BiologyExternal DOI
Issue
5Volume
27Page range
2055-2065Department affiliated with
- Biochemistry Publications
Full text available
- Yes
Peer reviewed?
- Yes
Legacy Posted Date
2018-08-14First Open Access (FOA) Date
2018-08-14First Compliant Deposit (FCD) Date
2018-08-14Usage metrics
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