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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\u20137). 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\u2019s 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\u201335% (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 \u03b1-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.<\/jats:p>","DOI":"10.1007\/s00249-019-01420-0","type":"journal-article","created":{"date-parts":[[2020,1,4]],"date-time":"2020-01-04T16:02:16Z","timestamp":1578153736000},"page":"145-153","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":21,"title":["Amyloid formation of bovine insulin is retarded in moderately acidic pH and by addition of short-chain alcohols"],"prefix":"10.1007","volume":"49","author":[{"given":"David","family":"Bernson","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Almedina","family":"Mecinovic","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Md Tuhin","family":"Abed","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fredrik","family":"Lim\u00e9","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Per","family":"Jageland","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Magnus","family":"Palml\u00f6f","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elin K.","family":"Esbj\u00f6rner","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2020,1,4]]},"reference":[{"key":"1420_CR1","doi-asserted-by":"publisher","first-page":"491","DOI":"10.1038\/224491a0","volume":"224","author":"MJ Adams","year":"1969","unstructured":"Adams MJ, Blundell TL, Dodson EJ, Dodson GG, Vijayan M, Baker EN, Harding MM, Hodgkin DC, Rimmer B, Sheat S (1969) Structure of rhombohedral 2 zinc insulin crystals. 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