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- W2085343308 abstract "A simple process of in vitro folding has been developed for the preparation of hirudin dimer. A variant of recombinant hirudin with Asp33 replaced by Cys was expressed in yeast and isolated by HPLC. Crude Cys33-hirudin contains heterogeneous products that are made of one species of primary sequence. They were together reduced/denatured, and allowed to re-fold in the sodium bicarbonate buffer (pH 8.3) alone. Active, homogeneous Cys33-hirudin monomer folded spontaneously with a first order rate constant of 0.05 +/- 0.01 min-1, followed by the oxidation of two Cys33 to produce the pure dimer. The folding yield was 90%. On an equal weight basis, both Cys33-hirudin monomer and the dimer exhibit thrombin inhibitory activity comparable to that of wild-type hirudin. Due to the presence of an extra cysteine, the folding of active hirudin monomer (formation of three native disulfides) was accelerated by at least 12-fold." @default.
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- W2085343308 date "1993-12-20" @default.
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- W2085343308 title "Production of disulfide-linked hirudin dimer by in vitro folding" @default.
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- W2085343308 doi "https://doi.org/10.1016/0014-5793(93)81607-2" @default.
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