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Selection of binders from phage displayed antibody libraries using the BIAcore(TM) biosensor

Malmborg, Ann-Christin LU ; Dueñas, Marta; Ohlin, Mats LU ; Söderlind, Eskil LU and Borrebaeck, C. A K LU (1996) In Journal of Immunological Methods 198(1). p.51-57
Abstract

In this report we show that phage displayed antibodies can be selected based on dissociation rate constants, using a BIAcore(TM) biosensor. To demonstrate the principle, two Fab phage stocks displaying antibodies specific for hen egg lysozyme or phenyloxazolone were mixed in a ratio of 1:10 and injected over the biosensor chip containing immobilized lysozyme. Antigen-specific bound phages were eluted and analysed for specificity and phage titer. This procedure enriched for phages carrying specific antibodies. Selection of high affinity binders from phage libraries was then demonstrated with the BIAcore(TM) when phages were eluted and collected at different time points. Soluble antibody fragments were subsequently expressed and their... (More)

In this report we show that phage displayed antibodies can be selected based on dissociation rate constants, using a BIAcore(TM) biosensor. To demonstrate the principle, two Fab phage stocks displaying antibodies specific for hen egg lysozyme or phenyloxazolone were mixed in a ratio of 1:10 and injected over the biosensor chip containing immobilized lysozyme. Antigen-specific bound phages were eluted and analysed for specificity and phage titer. This procedure enriched for phages carrying specific antibodies. Selection of high affinity binders from phage libraries was then demonstrated with the BIAcore(TM) when phages were eluted and collected at different time points. Soluble antibody fragments were subsequently expressed and their kinetic parameters were determined. The time of elution was directly proportional to the affinity, due to decreased dissociation rate constants. This procedure offers a rapid and simple approach for selecting binders from phage libraries differing in antibody dissociation rate constants.

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author
organization
publishing date
type
Contribution to journal
publication status
published
keywords
BIAcore(TM), Dissociation rate constant, Phage display, Selection
in
Journal of Immunological Methods
volume
198
issue
1
pages
7 pages
publisher
Elsevier
external identifiers
  • Scopus:0029914097
ISSN
0022-1759
DOI
10.1016/0022-1759(96)00159-7
language
English
LU publication?
yes
id
aeabe1a9-b7c9-4271-8e9e-ce8c179dd5ac
date added to LUP
2016-04-19 14:08:48
date last changed
2016-04-22 13:26:58
@misc{aeabe1a9-b7c9-4271-8e9e-ce8c179dd5ac,
  abstract     = {<p>In this report we show that phage displayed antibodies can be selected based on dissociation rate constants, using a BIAcore(TM) biosensor. To demonstrate the principle, two Fab phage stocks displaying antibodies specific for hen egg lysozyme or phenyloxazolone were mixed in a ratio of 1:10 and injected over the biosensor chip containing immobilized lysozyme. Antigen-specific bound phages were eluted and analysed for specificity and phage titer. This procedure enriched for phages carrying specific antibodies. Selection of high affinity binders from phage libraries was then demonstrated with the BIAcore(TM) when phages were eluted and collected at different time points. Soluble antibody fragments were subsequently expressed and their kinetic parameters were determined. The time of elution was directly proportional to the affinity, due to decreased dissociation rate constants. This procedure offers a rapid and simple approach for selecting binders from phage libraries differing in antibody dissociation rate constants.</p>},
  author       = {Malmborg, Ann-Christin and Dueñas, Marta and Ohlin, Mats and Söderlind, Eskil and Borrebaeck, C. A K},
  issn         = {0022-1759},
  keyword      = {BIAcore(TM),Dissociation rate constant,Phage display,Selection},
  language     = {eng},
  number       = {1},
  pages        = {51--57},
  publisher    = {ARRAY(0xa876740)},
  series       = {Journal of Immunological Methods},
  title        = {Selection of binders from phage displayed antibody libraries using the BIAcore(TM) biosensor},
  url          = {http://dx.doi.org/10.1016/0022-1759(96)00159-7},
  volume       = {198},
  year         = {1996},
}