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Production, characterization, and antigen specificity of recombinant 62-71-3, a candidate monoclonal antibody for rabies prophylaxis in humans

Both, L; van Dolleweerd, C; Wright, E; Banyard, AC; Bulmer-Thomas, B; Selden, D; Altmann, F; Fooks, AR; Ma, JK (2013) Production, characterization, and antigen specificity of recombinant 62-71-3, a candidate monoclonal antibody for rabies prophylaxis in humans. FASEB JOURNAL, 27 (5). 2055 - 2065. ISSN 0892-6638 https://doi.org/10.1096/fj.12-219964
SGUL Authors: Ma, Julian Van Dolleweerd, Craig John

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Abstract

Rabies 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.—Both, L., van Dolleweerd, C., Wright, E., Banyard, A. C., Bulmer-Thomas, B., Selden, D., Altmann, F., Fooks, A. R., Ma, J. K.-C. Production, characterization, and antigen specificity of recombinant 62-71-3, a candidate monoclonal antibody for rabies prophylaxis in humans.

Item Type: Article
Additional Information: This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/us/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Keywords: Amino Acid Sequence, Antibodies, Monoclonal, Antibodies, Viral, Antibody Specificity, Carbohydrate Sequence, Glycoproteins, Neutralization Tests, Post-Exposure Prophylaxis, Rabies Vaccines, Rabies virus, Recombinant Proteins, Single-Chain Antibodies, Tobacco, Science & Technology, Life Sciences & Biomedicine, Biochemistry & Molecular Biology, Biology, Cell Biology, Life Sciences & Biomedicine - Other Topics, plant biotechnology, molecular pharming, PEP, tobacco, VIRUS-NEUTRALIZING EPITOPE, POSTEXPOSURE PROPHYLAXIS, TRANSGENIC PLANTS, IMMUNE GLOBULIN, LINEAR EPITOPE, GLYCOPROTEIN, LYSSAVIRUSES, GLYCOSYLATION, VACCINE, PATHOGENICITY
SGUL Research Institute / Research Centre: Academic Structure > Infection and Immunity Research Institute (INII)
Journal or Publication Title: FASEB JOURNAL
ISSN: 0892-6638
Dates:
DateEvent
1 May 2013Published
Web of Science ID: WOS:000318226100025
Download EPMC Full text (HTML)
URI: https://openaccess.sgul.ac.uk/id/eprint/101604
Publisher's version: https://doi.org/10.1096/fj.12-219964

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