Light chain shuffling of a high affinity antibody results in a drift in epitope recognition.
Human polyclonal and monoclonal antibodies against pathogens and toxins are potentially useful in the treatment of various diseases. A number of human monoclonal antibodies with protective capacity in vitro have been established by conventional hybridoma technology. However, with the development of phage-display technology, the possibility of specifically tailoring antigen-binding properties has improved substantially. We show here that the reactivity of a high affinity, virus-neutralizing human antibody against the AD-2 epitope of cytomegalovirus gB can be modified by introducing other Vkappa sequences together with the original VH sequence. The fine specificity, as determined by the requirement of particular amino acid residues in the epitope, is shifted in these new antibody fragments. It was also evident that the VH/Vkappa pairing was not promiscuous, since antibody fragments selected by phage display retained light chain sequences very similar to the original hybridoma-derived light chain, proving that a high affinity interaction was very dependent on a co-operativity between both variable domains. These findings show that phage display technology might modify the binding properties of pre-existing, high affinity antibodies.
Ohlin
M
M
Department of Immunotechnology, Lund University, P.O. Box 7031, S-220 07 Lund, Sweden.
Owman
H
H
Mach
M
M
Borrebaeck
C A
CA
D000595:Amino Acid Sequence; D000914:Antibodies, Viral / Q000276:immunology*; D000915:Antibody Affinity; D000956:Antigens, Viral / Q000276:immunology*; D001483:Base Sequence; D003090:Coliphages; D003587:Cytomegalovirus / Q000276:immunology*; D017931:DNA Primers / Q000737:chemistry; D000939:Epitopes; D005803:Genes, Immunoglobulin; D006801:Humans; D007145:Immunoglobulin kappa-Chains / Q000737:chemistry; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D011994:Recombinant Proteins / Q000737:chemistry
D000914:Antibodies, Viral; D000956:Antigens, Viral; D017931:DNA Primers; D000939:Epitopes; D007145:Immunoglobulin kappa-Chains; D011994:Recombinant Proteins