C - Chemistry – Metallurgy – 07 – K
Patent
C - Chemistry, Metallurgy
07
K
C07K 1/107 (2006.01) C07K 1/00 (2006.01) C07K 1/12 (2006.01) C07K 5/11 (2006.01) C07K 7/18 (2006.01) C07K 14/47 (2006.01) C07K 14/475 (2006.01) C12N 15/62 (2006.01)
Patent
CA 2140002
The invention provides for a chemical method for preparing a recombinant single copy polypeptide or a portion thereof with a modified terminal amino acid .alpha.-carbon reactive group selected from the group consisting of N-terminal .alpha.-amine, C-termi- nal .alpha.-carboxyl, and a combination thereof. The steps of the method involve forming the recombinant single copy polypeptide or a portion thereof so that the single copy polypeptide is protected with one or more biologically added protecting groups at the N- terminal .alpha.-amine, C-terminal .alpha.-carboxyl. The recombinant single copy polypeptide can then be reacted with up to three chemical protecting agents to selectively protect reactive side chain groups and thereby prevent side chain groups from being modified. The recombinant single copy polypeptide can be cleaved with at least one cleavage reagent specific for the biological protecting group to form an unprotected terminal amino acid .alpha.-carbon reactive group. The unprotected terminal amino acid .alpha.-carbon reactive group is modified with at least one chemical modifying agent. The side chain protected terminally modified single copy polypep- tide is then deprotected at the side chain groups to form a terminally modified recombinant single copy polypeptide. The number and sequence of steps in the method can be varied to achieve selective modification at the N- and/or C-terminal amino acid of a recombinantly produced polypeptide.
Coolidge Thomas R.
Holmquist Bart
Stout Jay
Wagner Fred W.
Bionebraska Inc.
Goudreau Gage Dubuc
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