Characterization of the epitope for anti-human respiratory syncytial virus F protein monoclonal antibody 101F using synthetic peptides and genetic approaches.
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.
Wu
Sheng-Jiun
SJ
Protein Engineering, Centocor R&D Inc., 145 King of Prussia Road, Radnor, PA 19087, USA.
Schmidt
Albert
A
Protein Engineering, Centocor R&D Inc., 145 King of Prussia Road, Radnor, PA 19087, USA.
Beil
Eric J
EJ
Protein Engineering, Centocor R&D Inc., 145 King of Prussia Road, Radnor, PA 19087, USA.
Day
Nicole D
ND
Immunobiology, Centocor R&D Inc., 145 King of Prussia Road, Radnor, PA 19087, USA.
Branigan
Patrick J
PJ
Immunobiology, Centocor R&D Inc., 145 King of Prussia Road, Radnor, PA 19087, USA.
Liu
Changbao
C
Immunobiology, Centocor R&D Inc., 145 King of Prussia Road, Radnor, PA 19087, USA.
Gutshall
Lester L
LL
Immunobiology, Centocor R&D Inc., 145 King of Prussia Road, Radnor, PA 19087, USA.
Palomo
Concepción
C
Centro Nacional de Microbiología, Instituto de Salud Carlos III, Majadahonda, 28220 Madrid, Spain.
Furze
Julie
J
Institute for Animal Health, Compton, Newbury, Berkshire RG20 7NN, UK.
Taylor
Geraldine
G
Institute for Animal Health, Compton, Newbury, Berkshire RG20 7NN, UK.
Melero
José A
JA
Centro Nacional de Microbiología, Instituto de Salud Carlos III, Majadahonda, 28220 Madrid, Spain.
Tsui
Ping
P
Molecular Discovery Technologies, Centocor R&D Inc., 145 King of Prussia Road, Radnor, PA 19087, USA.
Del Vecchio
Alfred M
AM
Immunobiology, Centocor R&D Inc., 145 King of Prussia Road, Radnor, PA 19087, USA.
Kruszynski
Marian
M
Protein Engineering, Centocor R&D Inc., 145 King of Prussia Road, Radnor, PA 19087, USA.
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
D000911:Antibodies, Monoclonal; D010446:Peptide Fragments; D014759:Viral Envelope Proteins; D014760:Viral Fusion Proteins; D014765:Viral Vaccines