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Nilsson, Anders
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Nilsson, A., Lindgren, J. & Eriksson Karlström, A. (2017). Intramolecular Thioether Crosslinking to Increase the Proteolytic Stability of Affibody Molecules. ChemBioChem (Print), 18(20), 2056-2062
Open this publication in new window or tab >>Intramolecular Thioether Crosslinking to Increase the Proteolytic Stability of Affibody Molecules
2017 (English)In: ChemBioChem (Print), ISSN 1439-4227, E-ISSN 1439-7633, Vol. 18, no 20, p. 2056-2062Article in journal (Refereed) Published
Abstract [en]

Protein therapeutics suffer from low oral bioavailability, mainly due to poor membrane permeability and digestion by gastrointestinal proteases. To improve proteolytic stability, intramolecular thioether crosslinks were introduced into a three-helix affibody molecule binding the human epidermal growth factor receptor (EGFR). Solid-phase peptide synthesis was used to produce an unmodified control protein domain and three different crosslinked protein domain variants: one with a thioether crosslink between the N-terminal lysine residue and a cysteine residue in the second loop region (denoted K4), a second with a crosslink between the C-terminal lysine residue and a cysteine residue in the first loop region (denoted K58), and a third with crosslinks in both positions (denoted K4K58). Circular dichroism (CD) and surface-plasmon-resonance-based (SPR-based) biosensor studies of the protein domains showed that the three-helix structure and high-affinity binding to EGFR were preserved in the crosslinked protein domains. In vitro digestion by gastrointestinal proteases demonstrated that the crosslinked protein domains showed increased stability towards pepsin and towards a combination of trypsin and chymotrypsin.

Place, publisher, year, edition, pages
WILEY-V C H VERLAG GMBH, 2017
Keywords
affibodies, crosslinking, protein engineering, protein modifications, thioethers
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:kth:diva-217433 (URN)10.1002/cbic.201700350 (DOI)000413353800015 ()28836374 (PubMedID)2-s2.0-85030102142 (Scopus ID)
Note

QC 20171117

Available from: 2017-11-17 Created: 2017-11-17 Last updated: 2017-11-17Bibliographically approved
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