Engineering, Expression in Transgenic Plants and Characterisation of E559, a Rabies Virus-Neutralising Monoclonal Antibody

Abstract

Rabies post-exposure prophylaxis (PEP) currently comprises administration of rabies vaccine together with rabies immunoglobulin (RIG) of either equine or human origin. In the developing world, RIG preparations are expensive, often in short supply, and of variable efficacy. Therefore, we are seeking to develop a monoclonal antibody cocktail to replace RIG. Here, we describe the cloning, engineering and production in plants of a candidate monoclonal antibody (E559) for inclusion in such a cocktail. The murine constant domains of E559 were replaced with human IgG1κ constant domains and the resulting chimeric mouse-human genes were cloned into plant expression vectors for stable nuclear transformation of Nicotiana tabacum. The plant-expressed, chimeric antibody was purified and biochemically characterized, was demonstrated to neutralize rabies virus in a fluorescent antibody virus neutralization assay, and conferred protection in a hamster challenge model.

Plant-based vaccines against viruses

“…Production and successful efficacy testing of anti-Rabies virus MAbs made in transgenic tobacco [147] High-yield production via rBeYDV of anti-Ebolavirus Zaire and West Nile virus MAbs [149,150] Transient expression via rTMV, and successful pre-and post-exposure efficacy trial in macaques, of anti-Ebola MAb cocktail [151,152] Successful efficacy trial in macaques of the ZMApp therapeutic MAb cocktail [153] Legend: rTMV = recombinant Tobacco mosaic virus, including ICON vectors. rBeYDV = recombinant Bean yellow dwarf mastrevirus.…”

“…As an object example, an important aspect of rabies disease prevention is therapy, given the many people in developing countries who get bitten by suspected rabid animals annuallyand a major development in this area is the production of potent rabies-neutralising antibodies in plants, given the prevailing situation of mainly equine-produced sera being in short supply and of variable quality. A consortium of researchers that includes South Africans have recently described the engineering and production in transgenic N tabacum plants of both a humanised IgG version of the broadly neutralising murine MAb E559, and the murine version [147]. Purification via agarose protein A/G affinity chromatography yielded 1.8 mg/kg biomass (0.04% TSP) for the chimaeric Mab, and 1.2 mg/kg biomass (0.03% TSP) for the murine Ab.…”

Emergence of Novel Coronavirus 2019-nCoV: Need for Rapid Vaccine and Biologics Development

“…Arabidopsis and Carrot [35] Human papilloma virus (HPV) HPV16-L1 Tobacco [36] HPVL1 VLPs Tomato [37] Influenza H3N2 Nucleoprotein Maize [38] Rabies G protein Maize [39] Tomato [40] Polio VP1 Tobacco and Lettuce [41] PV3 VLPs Tobacco [42] Ebola Ebola Immune Complex Tobacco [43] Antibodies Dengue E60 Tobacco [44] Human Immunodeficiency Virus (HIV) VRC01, P2G12 Tobacco [45,46] Rabies E559 Tobacco [47] 62-71-3 Tobacco [48] Single-chain antibody molecule (62-71-3) Tobacco [49] Ebola ZMapp Tobacco [50] Influenza KPF1-Antx Tobacco [51] Porcine epidemic diarrhea virus (PEDV) 2C10 Tobacco and Lettuce [52] Chikungunya CHKVmab Tobacco [53] West Nile Virus pE16 Tobacco [54] Many biopharmaceutical companies have shifted their momentum towards the plant expression system by knowing its importance. Various plants such as tobacco, duckweed, moss, alfalfa, and other plants have been used for the production of plant-made pharmaceuticals.…”

Antibodies from plants for bionanomaterials

“…Therapy and passive immunization CEA, 56,57 Human immunodeficiency virus, [58][59][60] Rabies virus, [61][62][63] Streptococcus mutans, 64,65 Ebola virus, 66 HSV, 67 ferritin, 68 interleukins, 69 EGFR, 70 CD40, 71 anthrax toxin, 72 Salmonella enterica lipopolysaccharide, 73 and EpCAM 74,75 from their intrinsic safety compared with other systems-they do not produce endotoxins like bacteria and they do not support the replication of animal viruses and prions like animal cells. The key features of plants for the production of recombinant antibodies are summarized in Figure 2.…”