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Apparent heterogeneity in the pIII-peptide fusion protein in single-phage clones isolated from peptide libraries

Noppe, Wim; Galaev, Igor LU ; Mattiasson, Bo LU and Deckmyn, Hans (2011) In Protein Engineering Design & Selection 24(9). p.721-726
Abstract
Ligand homogeneity is an important issue in affinity chromatography. Using phages expressing peptides on the pIII protein, a heterogeneity in the binding of monoclonal phages was observed during affinity chromatography on supermacroporous cryogels. Fractions with different apparent binding affinities could be separated by stepwise elution. When these different fractions were re-applied, the respective differences in affinity were retained. However, when phage fractions with different apparent affinities were first amplified, an offspring was generated with again variable affinities. As the sequence of the peptide insert was the same, the heterogeneity must be ascribed to differences in avidity and although no direct evidence could be... (More)
Ligand homogeneity is an important issue in affinity chromatography. Using phages expressing peptides on the pIII protein, a heterogeneity in the binding of monoclonal phages was observed during affinity chromatography on supermacroporous cryogels. Fractions with different apparent binding affinities could be separated by stepwise elution. When these different fractions were re-applied, the respective differences in affinity were retained. However, when phage fractions with different apparent affinities were first amplified, an offspring was generated with again variable affinities. As the sequence of the peptide insert was the same, the heterogeneity must be ascribed to differences in avidity and although no direct evidence could be generated, we hypothesize that this is possibly due to phages displaying different numbers of the same peptide as a consequence of either proteolytic or packaging events during the amplification step in Escherichia coli. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
supermacroporous cryogel, pIII fusion protein, heterogeneity
in
Protein Engineering Design & Selection
volume
24
issue
9
pages
721 - 726
publisher
Oxford University Press
external identifiers
  • wos:000294555600012
  • scopus:80052317897
ISSN
1741-0126
DOI
10.1093/protein/gzr033
language
English
LU publication?
yes
id
5df213af-faa7-4dde-8ecb-ddb4672d2187 (old id 2159101)
date added to LUP
2011-09-26 08:06:24
date last changed
2017-01-01 05:26:57
@article{5df213af-faa7-4dde-8ecb-ddb4672d2187,
  abstract     = {Ligand homogeneity is an important issue in affinity chromatography. Using phages expressing peptides on the pIII protein, a heterogeneity in the binding of monoclonal phages was observed during affinity chromatography on supermacroporous cryogels. Fractions with different apparent binding affinities could be separated by stepwise elution. When these different fractions were re-applied, the respective differences in affinity were retained. However, when phage fractions with different apparent affinities were first amplified, an offspring was generated with again variable affinities. As the sequence of the peptide insert was the same, the heterogeneity must be ascribed to differences in avidity and although no direct evidence could be generated, we hypothesize that this is possibly due to phages displaying different numbers of the same peptide as a consequence of either proteolytic or packaging events during the amplification step in Escherichia coli.},
  author       = {Noppe, Wim and Galaev, Igor and Mattiasson, Bo and Deckmyn, Hans},
  issn         = {1741-0126},
  keyword      = {supermacroporous cryogel,pIII fusion protein,heterogeneity},
  language     = {eng},
  number       = {9},
  pages        = {721--726},
  publisher    = {Oxford University Press},
  series       = {Protein Engineering Design & Selection},
  title        = {Apparent heterogeneity in the pIII-peptide fusion protein in single-phage clones isolated from peptide libraries},
  url          = {http://dx.doi.org/10.1093/protein/gzr033},
  volume       = {24},
  year         = {2011},
}