title: Stability Characterization of a Vaccine Antigen Based on the Respiratory Syncytial Virus Fusion Glycoprotein.
abstract: Infection with Respiratory Syncytial Virus (RSV) causes both upper and lower respiratory tract disease in humans, leading to significant morbidity and mortality in both young children and older adults. Currently, there is no licensed vaccine available, and therapeutic options are limited. During the infection process, the type I viral fusion (F) glycoprotein on the surface of the RSV particle rearranges from a metastable prefusion conformation to a highly stable postfusion form. In people naturally infected with RSV, most potent neutralizing antibodies are directed to the prefusion form of the F protein. Therefore, an engineered RSV F protein stabilized in the prefusion conformation (DS-Cav1) is an attractive vaccine candidate. Long-term stability at 4°C or higher is a desirable attribute for a commercial subunit vaccine antigen. To assess the stability of DS-Cav1, we developed assays using D25, an antibody which recognizes the prefusion F-specific antigenic site Ø, and a novel antibody 4D7, which was found to bind antigenic site I on the postfusion form of RSV F. Biophysical analysis indicated that, upon long-term storage at 4°C, DS-Cav1 undergoes a conformational change, adopting alternate structures that concomitantly lose the site Ø epitope and gain the ability to bind 4D7.
authors.affiliation: Infectious Diseases and Vaccines Discovery, MRL, Merck & Co., Inc., Kenilworth, New Jersey, United States of America.
authors.forename: Jessica A
authors.initials: JA
authors.lastname: Flynn
authors.affiliation: Infectious Diseases and Vaccines Discovery, MRL, Merck & Co., Inc., Kenilworth, New Jersey, United States of America.
authors.forename: Eberhard
authors.initials: E
authors.lastname: Durr
authors.affiliation: Infectious Diseases and Vaccines Discovery, MRL, Merck & Co., Inc., Kenilworth, New Jersey, United States of America.
authors.forename: Ryan
authors.initials: R
authors.lastname: Swoyer
authors.affiliation: Infectious Diseases and Vaccines Discovery, MRL, Merck & Co., Inc., Kenilworth, New Jersey, United States of America.
authors.forename: Pedro J
authors.initials: PJ
authors.lastname: Cejas
authors.affiliation: Infectious Diseases and Vaccines Discovery, MRL, Merck & Co., Inc., Kenilworth, New Jersey, United States of America.
authors.forename: Melanie S
authors.initials: MS
authors.lastname: Horton
authors.affiliation: Infectious Diseases and Vaccines Discovery, MRL, Merck & Co., Inc., Kenilworth, New Jersey, United States of America.
authors.forename: Jennifer D
authors.initials: JD
authors.lastname: Galli
authors.affiliation: Eurofins Lancaster Laboratories Professional Scientific Services, Lancaster, Pennsylvania, United States of America.
authors.forename: Scott A
authors.initials: SA
authors.lastname: Cosmi
authors.affiliation: Infectious Diseases and Vaccines Discovery, MRL, Merck & Co., Inc., Kenilworth, New Jersey, United States of America.
authors.forename: Amy S
authors.initials: AS
authors.lastname: Espeseth
authors.affiliation: Infectious Diseases and Vaccines Discovery, MRL, Merck & Co., Inc., Kenilworth, New Jersey, United States of America.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Bett
authors.affiliation: Infectious Diseases and Vaccines Discovery, MRL, Merck & Co., Inc., Kenilworth, New Jersey, United States of America.
authors.forename: Lan
authors.initials: L
authors.lastname: Zhang
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism; D057134:Antibodies, Neutralizing / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D000937:Antigen-Antibody Reactions / Q000276:immunology; D000941:Antigens / Q000276:immunology* / Q000378:metabolism; D000939:Epitopes / Q000276:immunology; D057809:HEK293 Cells; D006801:Humans; D046529:Microscopy, Electron, Transmission; D055550:Protein Stability; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification; D018357:Respiratory Syncytial Virus Infections / Q000517:prevention & control; D022261:Respiratory Syncytial Virus Vaccines / Q000276:immunology; D018113:Respiratory Syncytial Virus, Human / Q000378:metabolism*; D020349:Surface Plasmon Resonance; D022223:Vaccines, Subunit / Q000276:immunology* / Q000378:metabolism; D014760:Viral Fusion Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D000941:Antigens; D000939:Epitopes; D011994:Recombinant Proteins; D022261:Respiratory Syncytial Virus Vaccines; D022223:Vaccines, Subunit; D014760:Viral Fusion Proteins