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Intramolecular Thio...
Intramolecular Thioether Crosslinking to Increase the Proteolytic Stability of Affibody Molecules
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- Nilsson, Anders (författare)
- KTH,Proteinteknologi
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- Lindgren, Joel, 1982- (författare)
- KTH,Proteinteknologi
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- Eriksson Karlström, Amelie (författare)
- KTH,Proteinteknologi
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(creator_code:org_t)
- 2017-09-19
- 2017
- Engelska.
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Ingår i: ChemBioChem. - : WILEY-V C H VERLAG GMBH. - 1439-4227 .- 1439-7633. ; 18:20, s. 2056-2062
- Relaterad länk:
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https://urn.kb.se/re...
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visa fler...
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https://doi.org/10.1...
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visa färre...
Abstract
Ämnesord
Stäng
- 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.
Ämnesord
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
Nyckelord
- affibodies
- crosslinking
- protein engineering
- protein modifications
- thioethers
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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