title: Montanide ISA 720 vaccines: quality control of emulsions, stability of formulated antigens, and comparative immunogenicity of vaccine formulations.
abstract: Montanide ISA 720 is an experimental adjuvant, formulated as water-in-oil emulsions, that induces high antibody titers in several animal species. It has been used in human vaccine trials with malaria and HIV vaccines. The heightened response is likely due, in part, to the formation of a depot at the injection site. However, post-formulation modifications were seen with seven proteins tested during storage of ISA 720 formulations at 37 degrees C for 1 week and two proteins stored longer at 4 degrees C. Potency studies in mice, in which the stored vaccines were diluted into placebo emulsions for appropriate dosing, indicated that this instability could lead to loss of immunogenicity in the post-injection depot, limiting the allowable storage time of preformed vaccines. We describe point-of-injection formulation for ISA 720 vaccines that meets the requirement for in vitro stability. For preformed vaccines, addition of glycine or glycylglycine prevented antigen modification on storage at 37 degrees C, providing a potential way of stabilizing antigen/ISA 720 formulations for in vitro storage and the post-injection depot.
authors.affiliation: Malaria Vaccine Development Branch, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 5640 Fishers Lane, Twinbrook I Room 1118, Rockville, MD 20852, USA. amiles@niaid.nih.gov
authors.forename: Aaron P
authors.initials: AP
authors.lastname: Miles
authors.forename: Holly A
authors.initials: HA
authors.lastname: McClellan
authors.forename: Kelly M
authors.initials: KM
authors.lastname: Rausch
authors.forename: Daming
authors.initials: D
authors.lastname: Zhu
authors.forename: Michael D
authors.initials: MD
authors.lastname: Whitmore
authors.forename: Sanjay
authors.initials: S
authors.lastname: Singh
authors.forename: Laura B
authors.initials: LB
authors.lastname: Martin
authors.forename: Yimin
authors.initials: Y
authors.lastname: Wu
authors.forename: Birgitte K
authors.initials: BK
authors.lastname: Giersing
authors.forename: Anthony W
authors.initials: AW
authors.lastname: Stowers
authors.forename: Carole A
authors.initials: CA
authors.lastname: Long
authors.forename: Allan
authors.initials: A
authors.lastname: Saul
mesh_terms: D000276:Adjuvants, Immunologic / Q000008:administration & dosage / Q000592:standards*; D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood; D000941:Antigens / Q000008:administration & dosage / Q000737:chemistry / Q000276:immunology*; D000953:Antigens, Protozoan / Q000276:immunology; D004355:Drug Stability; D004356:Drug Storage; D004655:Emulsions; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D008353:Mannitol / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000276:immunology / Q000592:standards; D051379:Mice; D008807:Mice, Inbred BALB C; D023421:Models, Animal; D009829:Oleic Acids / Q000008:administration & dosage / Q000276:immunology / Q000592:standards*; D011786:Quality Control; D011994:Recombinant Proteins; D013696:Temperature; D014612:Vaccines / Q000008:administration & dosage / Q000737:chemistry* / Q000276:immunology*
chemical_list: D000276:Adjuvants, Immunologic; D000913:Antibodies, Protozoan; D000941:Antigens; D000953:Antigens, Protozoan; D004655:Emulsions; D009829:Oleic Acids; D011994:Recombinant Proteins; D014612:Vaccines; C001681:mannide monooleate; D008353:Mannitol