Neutralising human recombinant antibodies to human cytomegalovirus glycoproteins gB and gH.
A phage antibody display library of single chain fragment variable (scFv) was applied to develop anti-HCMV glycoprotein B (gB) and glycoprotein H (gH) neutralising libraries. To enrich for specific scFvs, the phage antibody was panned against cytomegalovirus epitopes derived from the N-terminal part of gB, the C-terminal part of gB and the N-terminal part of gH (NETIYNTTLKYGDV, VTSGSTKD and AASEALDPHAFHLLLNTYGR). A number of clones were differentiated by Bst N1 fingerprinting. After isolation of specific clones against each peptide, the neutralising effect of each clone was assessed by plaque reduction assay. This resulted in the isolation of eight neutralising scFv antibodies with 51-63% neutralising effects. Sequence analysis of three neutralising clones revealed the amino acids specificity changes in heavy and light chains of antibody molecules.
Nejatollahi
Foroogh
F
Department of Medical Microbiology, Manchester University, Manchester Royal Infirmary, 2nd Floor, Clinical Sciences Building, Oxford Road, Manchester M13 9WL, UK.
Hodgetts
Samantha J
SJ
Vallely
Pamela J
PJ
Burnie
James P
JP
D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000235:genetics* / Q000276:immunology*; D002478:Cells, Cultured; D003001:Cloning, Molecular; D003586:Cytomegalovirus Infections / Q000276:immunology; D003588:Cytopathogenic Effect, Viral / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000032:analysis; D009500:Neutralization Tests; D019151:Peptide Library; D010455:Peptides / Q000378:metabolism; D011994:Recombinant Proteins / Q000276:immunology; D012680:Sensitivity and Specificity; D014759:Viral Envelope Proteins / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D010455:Peptides; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C040495:glycoprotein B, Simplexvirus; C071440:glycoprotein H, Cytomegalovirus; C499358:glycoprotein H, Human cytomegalovirus