Back to results
US 7,803,763 B2Granted
Method of purifying preproinsulin
AssigneeSanofi-Aventis Deutschland GmbH
InventorsHorst Thurow, Hans Blumenstock, Chantalle Havenith
Life of this patent
Estimating the term…
Abstract
The invention relates to a method for the chromatographic purification of preproinsulins, in which higher molecular weight substances are removed from an aqueous solution of preproinsulin by a first chromatography on an anion exchanger in flow-through mode and a subsequent second chromatography on a cation exchanger in adsorption mode, and to a method for preparing insulins, which includes the method for preparing preproinsulins.
Claims4
A method for the chromatographic purification of preproinsulin of the formula 1,
- wherein
- X a) is a genetically encodable amino acid residue or
- b) is a peptide having from 2 to 35 amino acid residues, which starts and ends with in each case a basic amino acid residue, in particular Arg, and which, if it consists of more than 3 amino acid residues, starts and ends with in each case two basic amino acid residues, in particular Arg and/or Lys,
- R1 a) is hydrogen,
- b) is a genetically encodable amino acid residue or
- c) is a peptide having from 2 to 15 amino acid residues,
- R2 is a genetically encodable amino acid residue, and
- the residues A1-A20 correspond to the amino acid sequence of the A chain of human insulin or of an insulin analog and the residues B1-B30 correspond to the amino acid sequence of the B chain of human insulin or of an insulin analog;
- wherein said method for chromatographic purification of preproinsulin comprises:
- removing higher molecular weight substances from an aqueous solution of said preproinsulin by means of a first chromatography on an anion exchanger in flow-through mode and a subsequent second chromatography on a cation exchanger in adsorption mode and
- wherein said preproinsulin of formula (l) is consisting of the following amino acid sequence:
- (SEQ ID NO:
- 3)
- Ala-Thr-Thr-Ser-Thr-Gly-Asn-Ser-Ala-Arg-Phe-Val-
- Asn-Gln-His-Leu-Cys-Gly-Ser-His-Leu-Val-Glu-Ala-
- Leu-Tyr-Leu-Val-Cys-Gly-Glu-Arg-Gly-Phe-Phe-Tyr-
- Thr-Pro-Lys-Thr-Arg-Arg-Glu-Ala-Glu-Asp-Pro-Gln-
- Val-Gly-Gln-Val-Glu-Leu-Gly-Gly-Gly-Pro-Gly-Ala-
- Gly-Ser-Leu-Gln-Pro-Leu-Ala-Leu-Glu-Gly-Ser-Leu-
- Gln-Lys-Arg-Gly-Ile-Val-Glu-Gln-Cys-Cys-Thr-Ser-
- Ile-Cys-Ser-Leu-Tyr-Gln-Leu-Glu-Asn-Tyr-Cys-Gly
- and wherein
- X is a peptide chain having 35 amino acid residues with the sequence of simian C peptide,
- R1 is a peptide chain having 10 amino acid residues with the sequence Ala-Thr-Thr-Ser-Thr-Gly-Asn-Ser-Ala-Arg (SEQ ID NO:
- 5),
- R2 is the amino acid residue Gly,
- A1-A20 is a peptide chain with the sequence (only A1 to A20) of the A chain of human insulin,
- B1-B30 is a peptide chain having the sequence of the B chain of human insulin.
25 more elements
Dependencies are inferred from the claim text.