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Light chain shuffling of a high affinity antibody results in a drift in epitope recognition

Ohlin, Mats LU ; Owman, Henrik LU ; Mach, Michael and Borrebaeck, Carl LU (1996) In Molecular Immunology 33(1). p.47-56
Abstract
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... (More)
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. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
cytomegalovirus, human monoclonal antibody, glycoprotein B (gB), phage-display technology, antibody specificity
in
Molecular Immunology
volume
33
issue
1
pages
47 - 56
publisher
Pergamon
external identifiers
  • pmid:8604223
  • scopus:0029981792
ISSN
1872-9142
DOI
10.1016/0161-5890(95)00123-9
language
English
LU publication?
yes
id
c8596726-4a88-4b96-98c6-84ad97e4988c (old id 1110672)
date added to LUP
2008-07-28 10:12:10
date last changed
2017-07-02 04:20:01
@article{c8596726-4a88-4b96-98c6-84ad97e4988c,
  abstract     = {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.},
  author       = {Ohlin, Mats and Owman, Henrik and Mach, Michael and Borrebaeck, Carl},
  issn         = {1872-9142},
  keyword      = {cytomegalovirus,human monoclonal antibody,glycoprotein B (gB),phage-display technology,antibody specificity},
  language     = {eng},
  number       = {1},
  pages        = {47--56},
  publisher    = {Pergamon},
  series       = {Molecular Immunology},
  title        = {Light chain shuffling of a high affinity antibody results in a drift in epitope recognition},
  url          = {http://dx.doi.org/10.1016/0161-5890(95)00123-9},
  volume       = {33},
  year         = {1996},
}