title: Production of full-length soluble Plasmodium falciparum RH5 protein vaccine using a Drosophila melanogaster Schneider 2 stable cell line system.
abstract: The Plasmodium falciparum reticulocyte-binding protein homolog 5 (PfRH5) has recently emerged as a leading candidate antigen against the blood-stage human malaria parasite. However it has proved challenging to identify a heterologous expression platform that can produce a soluble protein-based vaccine in a manner compliant with current Good Manufacturing Practice (cGMP). Here we report the production of full-length PfRH5 protein using a cGMP-compliant platform called ExpreS(2), based on a Drosophila melanogaster Schneider 2 (S2) stable cell line system. Five sequence variants of PfRH5 were expressed that differed in terms of mutagenesis strategies to remove potential N-linked glycans. All variants bound the PfRH5 receptor basigin and were recognized by a panel of monoclonal antibodies. Analysis following immunization of rabbits identified quantitative and qualitative differences in terms of the functional IgG antibody response against the P. falciparum parasite. The antibodies induced by one protein variant were shown to be qualitatively similar to responses induced by other vaccine platforms. This work identifies Drosophila S2 cells as a clinically-relevant platform suited for the production of 'difficult-to-make' proteins from Plasmodium parasites, and identifies a PfRH5 sequence variant that can be used for clinical production of a non-glycosylated, soluble full-length protein vaccine immunogen.
authors.affiliation: The Jenner Institute, University of Oxford, Old Road Campus Research Building, Oxford, OX3 7DQ, UK.
authors.forename: Kathryn A
authors.initials: KA
authors.lastname: Hjerrild
authors.affiliation: The Jenner Institute, University of Oxford, Old Road Campus Research Building, Oxford, OX3 7DQ, UK.
authors.forename: Jing
authors.initials: J
authors.lastname: Jin
authors.affiliation: Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX1 3QU, UK.
authors.forename: Katherine E
authors.initials: KE
authors.lastname: Wright
authors.affiliation: The Jenner Institute, University of Oxford, Old Road Campus Research Building, Oxford, OX3 7DQ, UK.
authors.forename: Rebecca E
authors.initials: RE
authors.lastname: Brown
authors.affiliation: The Jenner Institute, University of Oxford, Old Road Campus Research Building, Oxford, OX3 7DQ, UK.
authors.forename: Jennifer M
authors.initials: JM
authors.lastname: Marshall
authors.affiliation: The Jenner Institute, University of Oxford, Old Road Campus Research Building, Oxford, OX3 7DQ, UK.
authors.forename: Geneviève M
authors.initials: GM
authors.lastname: Labbé
authors.affiliation: The Jenner Institute, University of Oxford, Old Road Campus Research Building, Oxford, OX3 7DQ, UK.
authors.forename: Sarah E
authors.initials: SE
authors.lastname: Silk
authors.affiliation: The Jenner Institute, University of Oxford, Old Road Campus Research Building, Oxford, OX3 7DQ, UK.
authors.forename: Catherine J
authors.initials: CJ
authors.lastname: Cherry
authors.affiliation: ExpreS2ion Biotechnologies, SCION-DTU Science Park, Agern Allé 1, Hørsholm DK-2970, Denmark.
authors.forename: Stine B
authors.initials: SB
authors.lastname: Clemmensen
authors.affiliation: ExpreS2ion Biotechnologies, SCION-DTU Science Park, Agern Allé 1, Hørsholm DK-2970, Denmark.
authors.forename: Thomas
authors.initials: T
authors.lastname: Jørgensen
authors.affiliation: The Jenner Institute, University of Oxford, Old Road Campus Research Building, Oxford, OX3 7DQ, UK.
authors.forename: Joseph J
authors.initials: JJ
authors.lastname: Illingworth
authors.affiliation: The Jenner Institute, University of Oxford, Old Road Campus Research Building, Oxford, OX3 7DQ, UK.
authors.forename: Daniel G W
authors.initials: DG
authors.lastname: Alanine
authors.affiliation: The Jenner Institute, University of Oxford, Old Road Campus Research Building, Oxford, OX3 7DQ, UK.
authors.forename: Kathryn H
authors.initials: KH
authors.lastname: Milne
authors.affiliation: The Jenner Institute, University of Oxford, Old Road Campus Research Building, Oxford, OX3 7DQ, UK.
authors.forename: Rebecca
authors.initials: R
authors.lastname: Ashfield
authors.affiliation: ExpreS2ion Biotechnologies, SCION-DTU Science Park, Agern Allé 1, Hørsholm DK-2970, Denmark.
authors.forename: Willem A
authors.initials: WA
authors.lastname: de Jongh
authors.affiliation: The Jenner Institute, University of Oxford, Old Road Campus Research Building, Oxford, OX3 7DQ, UK.
authors.forename: Alexander D
authors.initials: AD
authors.lastname: Douglas
authors.affiliation: Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX1 3QU, UK.
authors.forename: Matthew K
authors.initials: MK
authors.lastname: Higgins
authors.affiliation: The Jenner Institute, University of Oxford, Old Road Campus Research Building, Oxford, OX3 7DQ, UK.
authors.forename: Simon J
authors.initials: SJ
authors.lastname: Draper
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D051926:Basigin / Q000276:immunology; D002352:Carrier Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D002460:Cell Line; D004331:Drosophila melanogaster; D007074:Immunoglobulin G / Q000276:immunology; D017780:Malaria Vaccines / Q000235:genetics / Q000276:immunology*; D009154:Mutation; D010963:Plasmodium falciparum / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D002352:Carrier Proteins; D007074:Immunoglobulin G; D017780:Malaria Vaccines; C531020:RH5 protein, Plasmodium falciparum; D051926:Basigin