title: Efficient In Vitro and In Vivo Activity of Glyco-Engineered Plant-Produced Rabies Monoclonal Antibodies E559 and 62-71-3.
abstract: Rabies is a neglected zoonotic disease that has no effective treatment after onset of illness. However the disease can be prevented effectively by prompt administration of post exposure prophylaxis which includes administration of passive immunizing antibodies (Rabies Immune Globulin, RIG). Currently, human RIG suffers from many restrictions including limited availability, batch-to batch inconsistencies and potential for contamination with blood-borne pathogens. Anti-rabies monoclonal antibodies (mAbs) have been identified as a promising alternative to RIG. Here, we applied a plant-based transient expression system to achieve rapid, high level production and efficacy of the two highly potent anti-rabies mAbs E559 and 62-71-3. Expression levels of up to 490 mg/kg of recombinant mAbs were obtained in Nicotiana benthamiana glycosylation mutants by using a viral based transient expression system. The plant-made E559 and 62-71-3, carrying human-type fucose-free N-glycans, assembled properly and were structurally sound as determined by mass spectrometry and calorimetric density measurements. Both mAbs efficiently neutralised diverse rabies virus variants in vitro. Importantly, E559 and 62-71-3 exhibited enhanced protection against rabies virus compared to human RIG in a hamster model post-exposure challenge trial. Collectively, our results provide the basis for the development of a multi-mAb based alternative to RIG.
authors.affiliation: Biosciences Unit, Council for Scientific and Industrial Research, Pretoria, South Africa.
authors.forename: Tsepo Lebiletsa
authors.initials: TL
authors.lastname: Tsekoa
authors.affiliation: Biosciences Unit, Council for Scientific and Industrial Research, Pretoria, South Africa.
authors.forename: Therese
authors.initials: T
authors.lastname: Lotter-Stark
authors.affiliation: Biosciences Unit, Council for Scientific and Industrial Research, Pretoria, South Africa.
authors.forename: Sindisiwe
authors.initials: S
authors.lastname: Buthelezi
authors.affiliation: Biosciences Unit, Council for Scientific and Industrial Research, Pretoria, South Africa.
authors.forename: Ereck
authors.initials: E
authors.lastname: Chakauya
authors.affiliation: Biosciences Unit, Council for Scientific and Industrial Research, Pretoria, South Africa.
authors.forename: Stoyan H
authors.initials: SH
authors.lastname: Stoychev
authors.affiliation: ARC-Onderstepoort Veterinary Institute, Onderstepoort, South Africa.
authors.forename: Claude
authors.initials: C
authors.lastname: Sabeta
authors.affiliation: ARC-Onderstepoort Veterinary Institute, Onderstepoort, South Africa.
authors.forename: Wonderful
authors.initials: W
authors.lastname: Shumba
authors.affiliation: ARC-Onderstepoort Veterinary Institute, Onderstepoort, South Africa.
authors.forename: Baby
authors.initials: B
authors.lastname: Phahladira
authors.affiliation: Kentucky Bioprocessing, Owensboro, Kentucky, United States.
authors.forename: Steve
authors.initials: S
authors.lastname: Hume
authors.affiliation: Kentucky Bioprocessing, Owensboro, Kentucky, United States.
authors.forename: Josh
authors.initials: J
authors.lastname: Morton
authors.affiliation: The Wistar Institute, Philadelphia, Pennsylvania, United States.
authors.forename: Charles E
authors.initials: CE
authors.lastname: Rupprecht
authors.affiliation: Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences, Vienna, Austria.
authors.forename: Herta
authors.initials: H
authors.lastname: Steinkellner
authors.affiliation: Mapp Biopharmaceutical, San Diego, California, United States.
authors.forename: Michael
authors.initials: M
authors.lastname: Pauly
authors.affiliation: Mapp Biopharmaceutical, San Diego, California, United States.
authors.forename: Larry
authors.initials: L
authors.lastname: Zeitlin
authors.affiliation: Mapp Biopharmaceutical, San Diego, California, United States.
authors.forename: Kevin
authors.initials: K
authors.lastname: Whaley
authors.affiliation: Biosciences Unit, Council for Scientific and Industrial Research, Pretoria, South Africa.
authors.forename: Rachel
authors.initials: R
authors.lastname: Chikwamba
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal / Q000008:administration & dosage / Q000096:biosynthesis* / Q000235:genetics; D000914:Antibodies, Viral / Q000008:administration & dosage / Q000096:biosynthesis* / Q000235:genetics; D003001:Cloning, Molecular; D005260:Female; D015870:Gene Expression; D005822:Genetic Vectors / Q000737:chemistry / Q000378:metabolism; D007116:Immunization, Passive; D008647:Mesocricetus; D009500:Neutralization Tests; D030821:Plants, Genetically Modified; D011818:Rabies / Q000276:immunology / Q000401:mortality / Q000517:prevention & control* / Q000821:virology; D011819:Rabies Vaccines / Q000008:administration & dosage / Q000096:biosynthesis; D011820:Rabies virus / Q000187:drug effects / Q000254:growth & development / Q000276:immunology / Q000472:pathogenicity; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000096:biosynthesis / Q000235:genetics; D016019:Survival Analysis; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D011819:Rabies Vaccines; D011994:Recombinant Proteins