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. In the case of circular dichroism (CD) spectroscopy, the same buffers was used, but with 10 mM Mops
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. Once protein unfolding reached completion in the first jump, the sample was cooled and incubated at 25°C for 1000 s, which was sufficient for completion of the helical refolding (as indicated by flattening of the Θ^sub 222^(t)) but not for the disk reconstitution (as indicated by DSC, EM, and gel electrophoresis)
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. Interestingly, a closer scrutiny of the insulin aggregates obtained after 1-min incubation at 60°C indicates that clusters of swollen insulin particles tend to align within the immediate vicinity of the early fibers (Fig
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