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Directed evolution of chemotaxis inhibitory protein of Staphylococcus aureus generates biologically functional variants with reduced interaction with human antibodies

Gustafsson, Erika LU ; Rosén, Anna; Barchan, Karin; Kessel, Kok P.M.; Haraldsson, Karin; Lindman, Stina LU ; Forsberg, Cecilia; Ljung, Lill; Bryder, Karin and Walse, Björn, et al. (2010) In Protein Engineering Design & Selection 23(2). p.91-101
Abstract
Chemotaxis inhibitory protein of Staphylococcus aureus (CHIPS) is a protein that binds and blocks the C5a receptor (C5aR) and formylated peptide receptor, thereby inhibiting the immune cell recruitment associated with inflammation. If CHIPS was less reactive with existing human antibodies, it would be a promising anti-inflammatory drug candidate. Therefore, we applied directed evolution and computational/rational design to the CHIPS gene in order to generate new CHIPS variants displaying lower interaction with human IgG, yet retaining biological function. The optimization was performed in four rounds: one round of random mutagenesis to add diversity into the CHIPS gene and three rounds of DNA recombination by Fragment INduced Diversity... (More)
Chemotaxis inhibitory protein of Staphylococcus aureus (CHIPS) is a protein that binds and blocks the C5a receptor (C5aR) and formylated peptide receptor, thereby inhibiting the immune cell recruitment associated with inflammation. If CHIPS was less reactive with existing human antibodies, it would be a promising anti-inflammatory drug candidate. Therefore, we applied directed evolution and computational/rational design to the CHIPS gene in order to generate new CHIPS variants displaying lower interaction with human IgG, yet retaining biological function. The optimization was performed in four rounds: one round of random mutagenesis to add diversity into the CHIPS gene and three rounds of DNA recombination by Fragment INduced Diversity (FIND((R))). Every round was screened by phage selection and/or ELISA for decreased interaction with human IgG and retained C5aR binding. The mean binding of human anti-CHIPS IgG decreased with every round of evolution. For further optimization, new amino acid substitutions were introduced by rational design, based on the mutations identified during directed evolution. Finally, seven CHIPS variants with low interaction with human IgG and retained C5aR blocking capacity could be identified. (Less)
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publication status
published
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in
Protein Engineering Design & Selection
volume
23
issue
2
pages
91 - 101
publisher
Oxford University Press
external identifiers
  • pmid:19959567
  • wos:000274286200005
  • scopus:75649131490
ISSN
1741-0126
DOI
10.1093/protein/gzp062
language
English
LU publication?
yes
id
2e9d6a4e-51b0-431d-8c24-333c0c6cc2ba (old id 1516889)
date added to LUP
2010-01-04 11:53:08
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2018-05-29 09:38:03
@article{2e9d6a4e-51b0-431d-8c24-333c0c6cc2ba,
  abstract     = {Chemotaxis inhibitory protein of Staphylococcus aureus (CHIPS) is a protein that binds and blocks the C5a receptor (C5aR) and formylated peptide receptor, thereby inhibiting the immune cell recruitment associated with inflammation. If CHIPS was less reactive with existing human antibodies, it would be a promising anti-inflammatory drug candidate. Therefore, we applied directed evolution and computational/rational design to the CHIPS gene in order to generate new CHIPS variants displaying lower interaction with human IgG, yet retaining biological function. The optimization was performed in four rounds: one round of random mutagenesis to add diversity into the CHIPS gene and three rounds of DNA recombination by Fragment INduced Diversity (FIND((R))). Every round was screened by phage selection and/or ELISA for decreased interaction with human IgG and retained C5aR binding. The mean binding of human anti-CHIPS IgG decreased with every round of evolution. For further optimization, new amino acid substitutions were introduced by rational design, based on the mutations identified during directed evolution. Finally, seven CHIPS variants with low interaction with human IgG and retained C5aR blocking capacity could be identified.},
  author       = {Gustafsson, Erika and Rosén, Anna and Barchan, Karin and Kessel, Kok P.M. and Haraldsson, Karin and Lindman, Stina and Forsberg, Cecilia and Ljung, Lill and Bryder, Karin and Walse, Björn and Haas, Pieter-Jan and van Strijp, Jos A.G. and Furebring, Christina},
  issn         = {1741-0126},
  language     = {eng},
  number       = {2},
  pages        = {91--101},
  publisher    = {Oxford University Press},
  series       = {Protein Engineering Design & Selection},
  title        = {Directed evolution of chemotaxis inhibitory protein of Staphylococcus aureus generates biologically functional variants with reduced interaction with human antibodies},
  url          = {http://dx.doi.org/10.1093/protein/gzp062},
  volume       = {23},
  year         = {2010},
}