Tsekoa, Tsepo LLotter-Stark, ThereseButhelezi, Sindisiwe GChakauya, EreckStoychev, Stoyan HChikwamba, Rachel K2017-05-162017-05-162016-07Tsekoa TL, Lotter-Stark T, Buthelezi S, Chakauya E, Stoychev SH, Sabeta C, et al. 2016. Efficient In Vitro and In Vivo Activity of Glyco-Engineered Plant-Produced Rabies Monoclonal Antibodies E559 and 62-71-3. PLoS ONE 11(7): e0159313. https://doi.org/10.1371/journal.pone.01593131932-6203http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0159313doi: 10.1371/journal.pone.0159313http://hdl.handle.net/10204/9028Copyright: © 2016 Tsekoa et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.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 bloodborne 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 Nglycans, 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.enCC0 1.0 UniversalRabiesRabies immune globulinRIGRabies monoclonal antibodies E559Rabies monoclonal antibodies 62-71-3ArticleTsekoa, T. L., Lotter-Stark, T., Buthelezi, S. G., Chakauya, E., Stoychev, S. H., & Chikwamba, R. K. (2016). Efficient in vitro and in vivo activity of glyco-engineered plant-produced rabies monoclonal antibodies E559 and 62-71-3. http://hdl.handle.net/10204/9028Tsekoa, Tsepo L, Therese Lotter-Stark, Sindisiwe G Buthelezi, Ereck Chakauya, Stoyan H Stoychev, and Rachel K Chikwamba "Efficient in vitro and in vivo activity of glyco-engineered plant-produced rabies monoclonal antibodies E559 and 62-71-3." (2016) http://hdl.handle.net/10204/9028Tsekoa TL, Lotter-Stark T, Buthelezi SG, Chakauya E, Stoychev SH, Chikwamba RK. Efficient in vitro and in vivo activity of glyco-engineered plant-produced rabies monoclonal antibodies E559 and 62-71-3. 2016; http://hdl.handle.net/10204/9028.TY - Article AU - Tsekoa, Tsepo L AU - Lotter-Stark, Therese AU - Buthelezi, Sindisiwe G AU - Chakauya, Ereck AU - Stoychev, Stoyan H AU - Chikwamba, Rachel K AB - 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 bloodborne 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 Nglycans, 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. DA - 2016-07 DB - ResearchSpace DP - CSIR KW - Rabies KW - Rabies immune globulin KW - RIG KW - Rabies monoclonal antibodies E559 KW - Rabies monoclonal antibodies 62-71-3 LK - https://researchspace.csir.co.za PY - 2016 SM - 1932-6203 T1 - Efficient in vitro and in vivo activity of glyco-engineered plant-produced rabies monoclonal antibodies E559 and 62-71-3 TI - Efficient in vitro and in vivo activity of glyco-engineered plant-produced rabies monoclonal antibodies E559 and 62-71-3 UR - http://hdl.handle.net/10204/9028 ER -