Neutralising human recombinant antibodies to human cytomegalovirus glycoproteins gB and gH |
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Authors: | Nejatollahi Foroogh Hodgetts Samantha J Vallely Pamela J Burnie James P |
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Affiliation: | Department of Medical Microbiology, Manchester University, Manchester Royal Infirmary, 2nd Floor, Clinical Sciences Building, Oxford Road, Manchester M13 9WL, UK. |
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Abstract: | 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. |
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Keywords: | Cytomegalovirus Single chain fragment variable Therapy Neutralizing antibody |
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