title: 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.
authors: Research Centre for Infection and Immunity, Division of Clinical Sciences, St George's University of London, United Kingdom.
authors: Craig J
authors: CJ
authors: van Dolleweerd
authors: Research Centre for Infection and Immunity, Division of Clinical Sciences, St George's University of London, United Kingdom.
authors: Audrey Y-H
authors: AY
authors: Teh
authors: Wildlife Zoonoses and Vector Borne Disease Research Group, Animal Health and Veterinary Laboratories Agency (AHVLA), Surrey, United Kingdom.
authors: Ashley C
authors: AC
authors: Banyard
authors: Research Centre for Infection and Immunity, Division of Clinical Sciences, St George's University of London, United Kingdom.
authors: Leonard
authors: L
authors: Both
authors: Council for Scientific and Industrial Research (CSIR), Biosciences, Pretoria, South Africa.
authors: Hester C T
authors: HC
authors: Lotter-Stark
authors: Council for Scientific and Industrial Research (CSIR), Biosciences, Pretoria, South Africa.
authors: Tsepo
authors: T
authors: Tsekoa
authors: Agricultural Research Council-Onderstepoort Veterinary Institute (ARC-OVI), OIE Rabies Reference Laboratory, Onderstepoort, Pretoria, South Africa.
authors: Baby
authors: B
authors: Phahladira
authors: Agricultural Research Council-Onderstepoort Veterinary Institute (ARC-OVI), OIE Rabies Reference Laboratory, Onderstepoort, Pretoria, South Africa.
authors: Wonderful
authors: W
authors: Shumba
authors: Council for Scientific and Industrial Research (CSIR), Biosciences, Pretoria, South Africa.
authors: Ereck
authors: E
authors: Chakauya
authors: Agricultural Research Council-Onderstepoort Veterinary Institute (ARC-OVI), OIE Rabies Reference Laboratory, Onderstepoort, Pretoria, South Africa.
authors: Claude T
authors: CT
authors: Sabeta
authors: Department of Chemistry, University of Natural Resources and Life Sciences, Vienna, Austria.
authors: Clemens
authors: C
authors: Gruber
authors: Wildlife Zoonoses and Vector Borne Disease Research Group, Animal Health and Veterinary Laboratories Agency (AHVLA), Surrey, United Kingdom.
authors: Anthony R
authors: AR
authors: Fooks
authors: Council for Scientific and Industrial Research (CSIR), Biosciences, Pretoria, South Africa.
authors: Rachel K
authors: RK
authors: Chikwamba
authors: Research Centre for Infection and Immunity, Division of Clinical Sciences, St George's University of London, United Kingdom.
authors: Julian K-C
authors: JK
authors: Ma
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics / Q000627:therapeutic use*; D000914:Antibodies, Viral / Q000096:biosynthesis* / Q000235:genetics / Q000627:therapeutic use*; D006224:Cricetinae; D004195:Disease Models, Animal; D006801:Humans; D008647:Mesocricetus; D051379:Mice; D030821:Plants, Genetically Modified; D011818:Rabies / Q000517:prevention & control; D011820:Rabies virus / Q000276:immunology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000627:therapeutic use; D014026:Nicotiana / Q000235:genetics
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D011994:Recombinant Proteins
keywords: Nicotiana tabacum; RIG; monoclonal antibody; post-exposure prophylaxis; rabies