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US 4,216,141 AGranted
Method for cyclization of peptides
AssigneeThe Salk Institute for Biological Studies
InventorsJean E. F. Rivier, Robert F. Galyean
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AssigneeThe Salk Institute for Biological Studies
InventorsJean E. F. Rivier, Robert F. Galyean
Estimating the term…
Peptides containing a disulfide bond between two cysteine residues are formed by an oxidation process. The oxidation step is accomplished under conditions whereby the sulfhydryl concentration in a reaction mixture is maintained at substantially zero during the reaction. In the process, an aqueous solution of a peptide containing at least one pair of cysteine moities is added incrementally to a buffered solution containing an oxidizing agent. The period between addition of the increments of the peptide solution is sufficient that the reaction to form the disulfide bond occurs substantially instantaneously. The increments are of such size that the sulfhydryl concentration during the reaction remains substantially constant and at a level of substantially zero, equivalent to infinite dilution.
A method for forming a disulfide bridge in peptides containing at least one pair of cysteine moieties comprising preparing a solution of a peptide containing at least one pair of cysteine moieties, preparing a solution of an oxidizing agent, adding said peptide solution in increments to said solution of oxidizing agent, monitoring the pH of the reaction mixture comprising the solution of oxidizing agent and the increment of the peptide solution and maintaining the pH of the reaction mixture at a level of from about 6 to about 8, the period between addition and the size of said increments being such that the oxidizing reaction to form the disulfide bond occurs substantially instantaneously and the sulfhydryl concentration during the reaction remains substantially constant and at a level of substantially zero.
Dependencies are inferred from the claim text.