Leader Sequence for Higher Expression of Recombinant Proteins
AssigneeUnichem Laboratories Ltd
InventorsDhananjay Sathe, Sudeep Kumar, Sachin Prabhakar Bachate, Saikumar Kompelli, Rahul Subhash Chougule
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Abstract
The present invention relates to the leader sequence for higher expression of recombinant proteins. The invention further relates to the process for preparation of insulin and insulin analogues using leader sequence. The leader peptides significantly increase the expression of pre-proinsulin. The present invention also relates to the protein sequences prepared by fusion of fragments with the leader sequences of the present invention. The invention is demonstrated by preparing and Insulin and its analogues using said leader sequences.
Claims28
A process for producing insulin and insulin analogues comprising pre-proinsulin of Formula 1:
- R1—X1-X2-X3 Formula 1
- as an intermediate, wherein X1 is a ‘B’ chain of insulin or insulin analogues, X2 is a dipeptide selected from RR or KR or RK or KK, X3 is an ‘A’ chain of insulin or insulin analogues and R1 is a leader peptide sequence selected from
- a) a peptide having amino acid sequence as set forth of in SEQ ID NO:
- 2 or
- b) a peptide having at least 80% homology to a).
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A polypeptide comprising a leader peptide operably linked to the precursor of insulin or insulin analogues, wherein the leader peptide is selected from:
- a) a peptide having amino acid sequence as set forth in SEQ ID NO:
- 2,
- b) a peptide having at least 80% homology to a).
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A leader peptide sequence selected from:
- a) a peptide having amino acid sequence as set forth in SEQ ID NO:
- 2,
- b) a peptide having at least 80% homology to a).
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A nucleotide sequence encoding amino acid sequence of Formula 1, R1—X1-X2-X3, wherein X1 is a ‘B’ chain of insulin or insulin analogues, X2 is a dipeptide selected RR or KR or RK or KK, X3 is an ‘A’ chain of insulin or insulin analogues and R1 is selected from:
- a) a peptide having amino acid sequence as set forth in SEQ ID NO:
- 2,
- b) a peptide having at least 80% homology to a).
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Dependencies are inferred from the claim text.