Apparent heterogeneity in the pIII-peptide fusion protein in single-phage clones isolated from peptide libraries
(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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/2159101
- author
- Noppe, Wim ; Galaev, Igor LU ; Mattiasson, Bo LU and Deckmyn, Hans
- organization
- publishing date
- 2011
- 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
- pmid:21705774
- 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
- 2016-04-01 13:04:00
- date last changed
- 2022-01-27 17:06:34
@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}}, keywords = {{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}}, doi = {{10.1093/protein/gzr033}}, volume = {{24}}, year = {{2011}}, }