US2023072673A1PendingUtilityA1

C-peptides and proinsulin polypeptides comprising the same

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Assignee: ABSCI LLCPriority: Sep 25, 2018Filed: Jul 6, 2022Published: Mar 9, 2023
Est. expirySep 25, 2038(~12.2 yrs left)· nominal 20-yr term from priority
Inventors:Amy DoernerMichael CammarataTyler NuscaSuzanne A. SprungerMelissa Patterson
C07K 1/22C12Y 304/17002C07K 14/62C07K 14/435C07K 14/47C07K 1/145C12N 9/485C12N 9/96C07K 2319/00C12N 15/70
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Claims

Abstract

A connecting polypeptide has SEQ ID NO:73. A proinsulin polypeptide includes a mature insulin A-chain, a mature insulin B-chain, and a connecting peptide comprising SEQ ID NO: 73 linking the mature A-chain and the mature B-chain, wherein the connecting peptide is not a native human proinsulin C-peptide. The proinsulin polypeptides according to the invention can be made in high titers and in high purity.

Claims

exact text as granted — not AI-modified
What is claimed is: 
     
         1 . A connecting polypeptide comprising SEQ ID NO:73. 
     
     
         2 . A proinsulin polypeptide comprising:
 a mature insulin A-chain;   a mature insulin B-chain; and   a connecting peptide comprising SEQ ID NO:73 linking the mature A-chain and the mature B-chain, wherein the connecting peptide is not a native human proinsulin C-peptide.   
     
     
         3 . The proinsulin polypeptide of  claim 2 , wherein the mature insulin A-chain is selected from the group consisting of SEQ ID NO:1 and SEQ ID NO:6. 
     
     
         4 . The proinsulin polypeptide of  claim 3 , wherein the mature insulin A-chain comprises SEQ ID NO:6. 
     
     
         5 . The proinsulin polypeptide of  claim 2 , wherein the mature insulin B-chain is selected from the group consisting of SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:8, and SEQ ID NO:9. 
     
     
         6 . The proinsulin polypeptide of  claim 5 , wherein the mature insulin B-chain comprises SEQ ID NO:7. 
     
     
         7 . The proinsulin polypeptide of  claim 2 , wherein the proinsulin polypeptide lacks a signal peptide. 
     
     
         8 . The proinsulin polypeptide of  claim 2 , further comprising a carboxypeptidase B propeptide coupled to the N-terminal residue of the mature insulin B-chain, the carboxypeptidase B propeptide selected from the group consisting of SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, and SEQ ID NO:36. 
     
     
         9 . The proinsulin polypeptide of  claim 8 , wherein the carboxypeptidase B propeptide comprises SEQ ID NO:36. 
     
     
         10 . The proinsulin polypeptide of  claim 2 , further comprising a propeptide coupled to the N-terminal residue of the mature insulin B-chain, wherein the propeptide is SEQ ID NO:37. 
     
     
         11 . An expression construct comprising a polynucleotide sequence encoding the insulin polypeptide of  claim 2 . 
     
     
         12 . A host cell comprising the expression construct of  claim 11 . 
     
     
         13 . A proinsulin polypeptide comprising:
 a mature insulin A-chain comprising SEQ ID NO:6;   a mature insulin B-chain comprising SEQ ID NO:7; and   a connecting peptide comprising SEQ ID NO: 73 linking the mature A-chain and the mature B-chain, wherein the connecting peptide is not a native human proinsulin C-peptide.   
     
     
         14 . The proinsulin polypeptide of  claim 13 , wherein the proinsulin polypeptide lacks a signal peptide. 
     
     
         15 . The proinsulin polypeptide of  claim 13 , further comprising a carboxypeptidase B propeptide coupled to the N-terminal residue of the mature insulin B-chain, the carboxypeptidase B propeptide selected from the group consisting of SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, and SEQ ID NO:36. 
     
     
         16 . The proinsulin polypeptide of  claim 15 , wherein the carboxypeptidase B propeptide comprises SEQ ID NO:36. 
     
     
         17 . An expression construct comprising a polynucleotide sequence encoding the insulin polypeptide of  claim 13 . 
     
     
         18 . A host cell comprising the expression construct of  claim 17 . 
     
     
         19 . A proinsulin polypeptide comprising:
 a mature insulin A-chain comprising SEQ ID NO:6;   a mature insulin B-chain comprising SEQ ID NO:7;   a connecting peptide comprising SEQ ID NO:73 linking the mature A-chain and the mature B-chain, wherein the connecting peptide is not a native human proinsulin C-peptide; and   a carboxypeptidase B propeptide coupled to the N-terminal residue of the mature insulin B-chain, the carboxypeptidase B propeptide comprising SEQ ID NO:36.   
     
     
         20 . The proinsulin polypeptide of  claim 19 , wherein the proinsulin polypeptide lacks a signal peptide.

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