Multimeric humanized varicella-zoster virus antibody fragments to gH neutralize virus while monomeric fragments do not.
Murine monoclonal antibody 206 (MAb mu206) binds to gH, the varicella-zoster virus (VZV) fusogen, neutralizing the virus in vitro in the absence of complement and inhibiting cell-to-cell spread and egress of VZV in cultured cells. We have humanized this antibody to generate MAb hu206 by complementarity determining region grafting. MAb hu206 retained binding and in vitro neutralizing activity, as well as cross-reactivity with ten different VZV strains. Single-chain antibody fragments (scAb) derived from MAb hu206 were produced in Escherichia coli. These scAb retained the binding properties of the whole antibody. However, monomeric scAb exhibited markedly reduced neutralizing activity compared to the bivalent parental MAb hu206. Shortening the peptide linker joining the V(H) to the V(kappa) domain from 14 to 5 or even 0 residues encouraged multimerization and increased neutralizing efficacy. The fact that Fab fragments enzymatically generated from whole MAb hu206 lost their neutralizing potency lent support to the proposal that valency is important for VZV neutralization at this epitope.
Drew
P D
PD
University of Aberdeen, Department of Molecular and Cell Biology, Institute of Medical Sciences, Foresterhill, Aberdeen AB25 2ZD, UK1.
Moss
M T
MT
Scotgen Biopharmaceuticals Inc., Kettock Lodge, Aberdeen Science and Technology Park, Aberdeen AB22 8GU, UK3.
Pasieka
T J
TJ
Department of Microbiology, University of Iowa College of Medicine, Iowa City, IA 52242, USA2.
Grose
C
C
Department of Microbiology, University of Iowa College of Medicine, Iowa City, IA 52242, USA2.
Harris
W J
WJ
Scotgen Biopharmaceuticals Inc., Kettock Lodge, Aberdeen Science and Technology Park, Aberdeen AB22 8GU, UK3.|University of Aberdeen, Department of Molecular and Cell Biology, Institute of Medical Sciences, Foresterhill, Aberdeen AB25 2ZD, UK1.
Porter
A J R
AJR
University of Aberdeen, Department of Molecular and Cell Biology, Institute of Medical Sciences, Foresterhill, Aberdeen AB25 2ZD, UK1.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D002460:Cell Line; D004306:Dose-Response Relationship, Immunologic; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D006023:Glycoproteins / Q000276:immunology; D006562:Herpes Zoster / Q000276:immunology; D014645:Herpesvirus 3, Human / Q000737:chemistry / Q000276:immunology*; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D051379:Mice; D009500:Neutralization Tests; D015202:Protein Engineering; D014759:Viral Envelope Proteins / Q000276:immunology
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D006023:Glycoproteins; D007140:Immunoglobulin Fab Fragments; D014759:Viral Envelope Proteins