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Updated: May 21, 2026

Rapid Generation of Amyloid from Native Proteins In vitro
Published on: December 5, 2013
Regeneration of insulin monomers from amyloid fibrils by a NH3/H2O2 two-step method
Rui Liu1, Rongxin Su, Yanjun Yu
1State Key Laboratory of Chemical Engineering, School of Chemical Engineering and Technology, Tianjin University, 92 Weijin Road, Nankai District, Tianjin 300072, China.
Abstract:
We have developed a NH(3)/H(2)O(2) two-step method for the recovery of insulin monomers from amyloid fibrils by modulating the cleavage and regeneration of disulfide bonds. Insulin fibrils were disaggregated into insulin A- and B-chains in 14 M (w/v) NH(4)OH for 2 h at 60 °C. Insulin monomers, with a MW of ~5,882 Da, were then regenerated by oxidation of sulfhydryls with 30 % (w/v) H(2)O(2) (10 M) for 12 h at 25 °C. No two A-chains or two B-chains of insulin formed during the oxidation process. Because of the inconformity of the optimal reduction and oxidation temperature, the NH(3)/H(2)O(2) two-step method is more practical than the NH(3)/H(2)O(2) coupling method.
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