Abstract
The folding of biotechnological precursor of the human insulin precursor was carried out from solubilized inclusion bodies without a preliminary oxidizing or reducing its Cys residues. The inclusion bodies were dissolved in 8 M urea with the addition of 10 mM 2-mercaptoethanol. Hydrophobic cell components were removed from the solution by passing through a neutral weakly hydrophobic sorbent, the solution was five times diluted and refolded upon addition of 0.3 mM cystine for initiation of disulfide rearrangement. The presence of nucleic acids and cell protein impurities does not affect the folding efficiency. The resulting precursor of folded human insulin was purified by metal-chelate affinity chromatography and converted into insulin by two-stage enzymatic cleavage.
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Abbreviations
- HP:
-
hybrid protein
- sHP:
-
HP sulfonate
- rHP:
-
renatured HP with wild-type disulfide bonds in the proinsulin part of molecule
References
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For previous communication, see Protein Exp. Purif. 2002, vol. 26, pp. 187–193.
Original Russian Text © R.V. Tikhonov, A.N. Wulfson, M.P. Kirpichnikov, 2008, published in Bioorganicheskaya Khimiya, 2008, Vol. 34, No. 1, pp. 63–66.
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Tikhonov, R.V., Wulfson, A.N. & Kirpichnikov, M.P. Recombinant human insulin: X simplification of folding of the biotechnological precursor of recombinant human insulin. Russ J Bioorg Chem 34, 56–59 (2008). https://doi.org/10.1134/S106816200801007X
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DOI: https://doi.org/10.1134/S106816200801007X