title: Characterization of the epitope for anti-human respiratory syncytial virus F protein monoclonal antibody 101F using synthetic peptides and genetic approaches.
abstract: Chimeric 101F (ch101F) is a mouse-human chimeric anti-human respiratory syncytial virus (HRSV) neutralizing antibody that recognizes residues within antigenic site IV, V, VI of the fusion (F) glycoprotein. The binding of ch101F to a series of peptides overlapping aa 422-438 spanning antigenic site IV, V, VI was analysed. Residues 423-436 comprise the minimal peptide sequence for ch101F binding. Substitution analysis revealed that R429 and K433 are critical for ch101F binding, whilst K427 makes a minor contribution. Binding of ch101F to a series of single mutations at positions 427, 429 and 433 in the F protein expressed recombinantly on the cell surface confirmed the peptide results. Sequence analysis of viruses selected for resistance to neutralization by ch101F indicated that a single change (K433T) in the F protein allowed ch101F escape. The results confirm that ch101F and palivizumab have different epitope specificity and define key residues for ch101F recognition.
authors.affiliation: Protein Engineering, Centocor R&D Inc., 145 King of Prussia Road, Radnor, PA 19087, USA.
authors.forename: Sheng-Jiun
authors.initials: SJ
authors.lastname: Wu
authors.affiliation: Protein Engineering, Centocor R&D Inc., 145 King of Prussia Road, Radnor, PA 19087, USA.
authors.forename: Albert
authors.initials: A
authors.lastname: Schmidt
authors.affiliation: Protein Engineering, Centocor R&D Inc., 145 King of Prussia Road, Radnor, PA 19087, USA.
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Beil
authors.affiliation: Immunobiology, Centocor R&D Inc., 145 King of Prussia Road, Radnor, PA 19087, USA.
authors.forename: Nicole D
authors.initials: ND
authors.lastname: Day
authors.affiliation: Immunobiology, Centocor R&D Inc., 145 King of Prussia Road, Radnor, PA 19087, USA.
authors.forename: Patrick J
authors.initials: PJ
authors.lastname: Branigan
authors.affiliation: Immunobiology, Centocor R&D Inc., 145 King of Prussia Road, Radnor, PA 19087, USA.
authors.forename: Changbao
authors.initials: C
authors.lastname: Liu
authors.affiliation: Immunobiology, Centocor R&D Inc., 145 King of Prussia Road, Radnor, PA 19087, USA.
authors.forename: Lester L
authors.initials: LL
authors.lastname: Gutshall
authors.affiliation: Centro Nacional de Microbiología, Instituto de Salud Carlos III, Majadahonda, 28220 Madrid, Spain.
authors.forename: Concepción
authors.initials: C
authors.lastname: Palomo
authors.affiliation: Institute for Animal Health, Compton, Newbury, Berkshire RG20 7NN, UK.
authors.forename: Julie
authors.initials: J
authors.lastname: Furze
authors.affiliation: Institute for Animal Health, Compton, Newbury, Berkshire RG20 7NN, UK.
authors.forename: Geraldine
authors.initials: G
authors.lastname: Taylor
authors.affiliation: Centro Nacional de Microbiología, Instituto de Salud Carlos III, Majadahonda, 28220 Madrid, Spain.
authors.forename: José A
authors.initials: JA
authors.lastname: Melero
authors.affiliation: Molecular Discovery Technologies, Centocor R&D Inc., 145 King of Prussia Road, Radnor, PA 19087, USA.
authors.forename: Ping
authors.initials: P
authors.lastname: Tsui
authors.affiliation: Immunobiology, Centocor R&D Inc., 145 King of Prussia Road, Radnor, PA 19087, USA.
authors.forename: Alfred M
authors.initials: AM
authors.lastname: Del Vecchio
authors.affiliation: Protein Engineering, Centocor R&D Inc., 145 King of Prussia Road, Radnor, PA 19087, USA.
authors.forename: Marian
authors.initials: M
authors.lastname: Kruszynski
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D019921:Biotinylation; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D051379:Mice; D010446:Peptide Fragments / Q000276:immunology; D018357:Respiratory Syncytial Virus Infections / Q000276:immunology; D018113:Respiratory Syncytial Virus, Human / Q000235:genetics* / Q000276:immunology; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology; D014760:Viral Fusion Proteins / Q000235:genetics* / Q000276:immunology*; D014765:Viral Vaccines
chemical_list: D000911:Antibodies, Monoclonal; D010446:Peptide Fragments; D014759:Viral Envelope Proteins; D014760:Viral Fusion Proteins; D014765:Viral Vaccines