title: Large-scale purification and characterization of malaria vaccine candidate antigen Pvs25H for use in clinical trials.
abstract: The budding yeast Saccharomyces cerevisiae has been used to express the recombinant protein Pvs25H, currently the only candidate transmission-blocking vaccine against Plasmodium vivax malaria. This molecule contains four epidermal growth factor-like domains and is expressed as at least two stable monomeric forms with different physicochemical properties. Pvs25H-A is apparently homogeneous and seems to have a correct disulfide bond structure. By contrast, Pvs25H-B is produced as a heterogeneous population of molecules, some of which are associated with an as yet unidentified chromophore, and it contains both internal and N-terminal cleavages. We report here a procedure for successfully separating these two forms with a process suitable for clinical production of this antigen.
authors.affiliation: Malaria Vaccine Development Unit, National Institute of Allergy and Infectious Diseases, Rockville, Maryland 20852, USA.
authors.forename: Aaron P
authors.initials: AP
authors.lastname: Miles
authors.forename: Yanling
authors.initials: Y
authors.lastname: Zhang
authors.forename: Allan
authors.initials: A
authors.lastname: Saul
authors.forename: Anthony W
authors.initials: AW
authors.lastname: Stowers
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000913:Antibodies, Protozoan / Q000276:immunology; D000941:Antigens / Q000737:chemistry* / Q000302:isolation & purification*; D000953:Antigens, Protozoan / Q000737:chemistry* / Q000302:isolation & purification*; D000954:Antigens, Surface / Q000737:chemistry* / Q000302:isolation & purification*; D015153:Blotting, Western; D002845:Chromatography; D002851:Chromatography, High Pressure Liquid; D002986:Clinical Trials as Topic; D004220:Disulfides; D005285:Fermentation; D006801:Humans; D008288:Malaria / Q000517:prevention & control; D017780:Malaria Vaccines / Q000737:chemistry* / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D010966:Plasmodium vivax / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism*
chemical_list: D000913:Antibodies, Protozoan; D000941:Antigens; D000953:Antigens, Protozoan; D000954:Antigens, Surface; D004220:Disulfides; D017780:Malaria Vaccines; C121473:Pvs25 protein, P vivax; D011994:Recombinant Proteins