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A mild hydrophobic ...
A mild hydrophobic interaction chromatography involving polyethylene glycol immobilized to agarose media refolding recombinant Staphylococcus aureus elongation factor G
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Li, Jing-Jing (författare)
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- Venkataramana,, Musturi (författare)
- Uppsala universitet,Institutionen för cell- och molekylärbiologi
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Wang, Ai-Qing (författare)
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visa fler...
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- Sanyal, Suparna (författare)
- Uppsala universitet,Institutionen för cell- och molekylärbiologi
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- Janson, Jan-Christer (författare)
- Uppsala universitet,Institutionen för ytbioteknik med Centrum för ytbioteknik,Institutionen för fysikalisk och analytisk kemi
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Su, Zhi-Guo (författare)
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visa färre...
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(creator_code:org_t)
- 2005
- 2005
- Engelska.
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Ingår i: Protein Expression and Purification. ; 40:2, s. 327-335
- Relaterad länk:
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https://urn.kb.se/re...
Abstract
Ämnesord
Stäng
- Recombinant Staphylococcus aureus elongation factor G (EF-G) is difficult to refold by dilution due to the formation of large amounts of misfolded structures. However, refolding of EF-G by adsorption to a chromatographic column packed with immobilized polyethylene glycol 20,000 (PEG 20 K) followed by pulse elution with 8 M urea resulted in 88% mass recovery and 80% of correctly refolded structure. The PEG 20 K was coupled to brominated allyl group derivatized Sepharose High Performance to construct a mild hydrophobic adsorbent. Various other hydrophobic interaction adsorbents were also attempted to refold EF-G. However, ligands with high hydrophobicity tended to misfold EF-G, resulting in irreversible adsorption. Various solvents, detergents, and low temperature as well as 8 M urea were tried to release bound EF-G. Only pulse elution with 8 M urea was efficient. Urea concentrations favorable for efficiently refolding EF-G were investigated. Low urea concentration produced more misfolded structures.
Nyckelord
- Elongation factor G; Refolding
- Immobilized polyethylene glycol
- Hydrophobic interaction chromatography
- Pulse elution
- Urea
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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