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Amyloid formation o...
Amyloid formation of bovine insulin is retarded in moderately acidic pH and by addition of short-chain alcohols
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- Bernson, David, 1986 (författare)
- Chalmers tekniska högskola,Chalmers University of Technology
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- Basic, Almedina, 1992 (författare)
- Chalmers tekniska högskola,Chalmers University of Technology
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- Abed, MD Tuhin, 1991 (författare)
- Chalmers tekniska högskola,Chalmers University of Technology
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Lime, Fredrik (författare)
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Jageland, Per (författare)
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Palmlof, Magnus (författare)
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- Esbjörner Winters, Elin, 1978 (författare)
- Chalmers tekniska högskola,Chalmers University of Technology
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(creator_code:org_t)
- 2020-01-04
- 2020
- Engelska.
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Ingår i: European Biophysics Journal. - : Springer Science and Business Media LLC. - 1432-1017 .- 0175-7571. ; 49:2, s. 145-153
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Abstract
Ämnesord
Stäng
- Protein aggregation and amyloid formation are associated with multiple human diseases, but are also a problem in protein production. Understanding how aggregation can be modulated is therefore of importance in both medical and industrial contexts. We have used bovine insulin as a model protein to explore how amyloid formation is affected by buffer pH and by the addition of short-chain alcohols. We find that bovine insulin forms amyloid fibrils, albeit with different rates and resulting fibril morphologies, across a wide pH range (2-7). At pH 4.0, bovine insulin displayed relatively low aggregation propensity in combination with high solubility; this condition was therefore chosen as basis for further exploration of how bovine insulin's native state can be stabilized in the presence of short-chain alcohols that are relevant because of their common use as eluents in industrial-scale chromatography purification. We found that ethanol and isopropanol are efficient modulators of bovine insulin aggregation, providing a three to four times retardation of the aggregation kinetics at 30-35% (vol/vol) concentration; we attribute this to the formation of oligomers, which we detected by AFM. We discuss this effect in terms of reduced solvent polarity and show, by circular dichroism recordings, that a concomitant change in alpha-helical packing of the insulin monomer occurs in ethanol. Our results extend current knowledge of how insulin aggregates, and may, although bovine insulin serves as a simplistic model, provide insights into how buffers and additives can be fine-tuned in industrial production of proteins in general and pharmaceutical insulin in particular.
Ämnesord
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
- NATURVETENSKAP -- Biologi -- Biofysik (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biophysics (hsv//eng)
- TEKNIK OCH TEKNOLOGIER -- Industriell bioteknik -- Annan industriell bioteknik (hsv//swe)
- ENGINEERING AND TECHNOLOGY -- Industrial Biotechnology -- Other Industrial Biotechnology (hsv//eng)
Nyckelord
- Insulin
- Circular dichroism
- Aggregation
- Thioflavin-T
- Kinetics
- Amyloid
Publikations- och innehållstyp
- art (ämneskategori)
- ref (ämneskategori)
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