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Transiently binding antibody fragments against Lewis x and sialyl-Lewis x

Johansson, R ; Ohlin, Mats LU orcid ; Jansson, B and Ohlson, S (2006) In Journal of Immunological Methods 312(1-2). p.20-26
Abstract
Biomolecular recognition is often characterised by low affinity where many weak interactions work either alone or in concert, resulting in an inherent dynamic situation. For example the well-studied weak binding of cell-cell interactions is predominantly based on a range of carbohydrates that interact with numerous (protein) ligands. Finding appropriate binders to these carbohydrate structures may pave the way for new analytical strategies based on low affinity, and recombinant antibody technology is a promising approach to the development of such reagents. We have in the present study characterised two low affinity human single chain antibody fragments (scFv) by surface plasmon resonance for use in such applications. The two clones, LeX1... (More)
Biomolecular recognition is often characterised by low affinity where many weak interactions work either alone or in concert, resulting in an inherent dynamic situation. For example the well-studied weak binding of cell-cell interactions is predominantly based on a range of carbohydrates that interact with numerous (protein) ligands. Finding appropriate binders to these carbohydrate structures may pave the way for new analytical strategies based on low affinity, and recombinant antibody technology is a promising approach to the development of such reagents. We have in the present study characterised two low affinity human single chain antibody fragments (scFv) by surface plasmon resonance for use in such applications. The two clones, LeX1 and sLeX10, had been selected from a naive phage display library against Lewis x (Le(x)) and sialyl Le(x) (sLe(x)), respectively. Both LeX1 and sLeX10 showed low affinity, with K-D values of 3.5 +/- 0.7 x 10(-5) M for Le(x) and 2.6 +/- 0.7 x 10(-5) M for sLe(x), respectively. Kinetic studies revealed the scFvs to be associated with fast dissociation rates, with K-d values higher than 0.1 s(-1) for both LeX1 and sLeX10. Apart from the Lewis structures Le(x) and sLe(x), we investigated the conforinational isomers Lewis a and sialyl-Lewis a together with the monosaccharide units of the Lewis structures, and both scFvs showed high specificity for their respective carbohydrate. Taking these observations together we have demonstrated that scFv with fast reaction kinetics and low affinity have the necessary characteristics for further development as specific tools in analytical strategies, e.g. differentiation of cells based on the various configurations of carbohydrate epitopes. (c) 2006 Elsevier B.V. All rights reserved. (Less)
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author
; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
surface plasmon, low affinity, sialyl-Lewis x, single chain Fv, Lewis x, resonance
in
Journal of Immunological Methods
volume
312
issue
1-2
pages
20 - 26
publisher
Elsevier
external identifiers
  • wos:000238605000003
  • pmid:16581086
  • scopus:33646903696
ISSN
1872-7905
DOI
10.1016/j.jim.2006.02.010
language
English
LU publication?
yes
id
eac00216-ac4c-4aef-bfe6-39600b62e590 (old id 404891)
date added to LUP
2016-04-01 15:50:04
date last changed
2021-02-17 08:12:01
@article{eac00216-ac4c-4aef-bfe6-39600b62e590,
  abstract     = {Biomolecular recognition is often characterised by low affinity where many weak interactions work either alone or in concert, resulting in an inherent dynamic situation. For example the well-studied weak binding of cell-cell interactions is predominantly based on a range of carbohydrates that interact with numerous (protein) ligands. Finding appropriate binders to these carbohydrate structures may pave the way for new analytical strategies based on low affinity, and recombinant antibody technology is a promising approach to the development of such reagents. We have in the present study characterised two low affinity human single chain antibody fragments (scFv) by surface plasmon resonance for use in such applications. The two clones, LeX1 and sLeX10, had been selected from a naive phage display library against Lewis x (Le(x)) and sialyl Le(x) (sLe(x)), respectively. Both LeX1 and sLeX10 showed low affinity, with K-D values of 3.5 +/- 0.7 x 10(-5) M for Le(x) and 2.6 +/- 0.7 x 10(-5) M for sLe(x), respectively. Kinetic studies revealed the scFvs to be associated with fast dissociation rates, with K-d values higher than 0.1 s(-1) for both LeX1 and sLeX10. Apart from the Lewis structures Le(x) and sLe(x), we investigated the conforinational isomers Lewis a and sialyl-Lewis a together with the monosaccharide units of the Lewis structures, and both scFvs showed high specificity for their respective carbohydrate. Taking these observations together we have demonstrated that scFv with fast reaction kinetics and low affinity have the necessary characteristics for further development as specific tools in analytical strategies, e.g. differentiation of cells based on the various configurations of carbohydrate epitopes. (c) 2006 Elsevier B.V. All rights reserved.},
  author       = {Johansson, R and Ohlin, Mats and Jansson, B and Ohlson, S},
  issn         = {1872-7905},
  language     = {eng},
  number       = {1-2},
  pages        = {20--26},
  publisher    = {Elsevier},
  series       = {Journal of Immunological Methods},
  title        = {Transiently binding antibody fragments against Lewis x and sialyl-Lewis x},
  url          = {http://dx.doi.org/10.1016/j.jim.2006.02.010},
  doi          = {10.1016/j.jim.2006.02.010},
  volume       = {312},
  year         = {2006},
}