Patent ID: 6051399
Filing Date: 2000-04-18
Classification: C07K,C12N,C12P

Abstract:
A method of producing a peptide having a C-terminal .alpha.-carboxamide comprising:converting a recombinant protein construct to a product peptide having a C-terminal .alpha.-carboxamide by contact of the recombinant protein construct with (a) an S-cyanylating agent and an amine compound, and (b) a CS1 cleaving agent; wherein,the recombinant protein construct has an amino acid sequence of the formula: ##STR1## -CS1- is a chemical cleavage site selected from the group consisting of AspGly, AspPro, AsnGly, Met and Trp;the CS1 cleaving agent is low pH for AspGly and AspPro, hydroxylanine for AsnGly, cyanogen bromide for Met, and BNPS-skatole or other oxidizing agent for Trp;-(Ln1)- is a linking peptide other than CS1, Cys or TargP, and is a spacer unit;-TargP- is a target peptide which is free of unblocked cysteine residues;n is 0 or 1;r is an integer from 1 to about 150;Yyy- is a leader group and includes a cleavage site at its C end;-Xxx is a tail group; andsteps (a) and (b) are carried out either in an order--a followed by b or an order--b followed by a.