N-terminal derivatisation of proteins with polysaccharides
First Claim
1. A method for producing a composition comprising a population of polysialic acid derivatives of a protein, wherein the derivatives comprise between 2 and 200 sialic acid units and wherein the population consists substantially only of N-terminal derivatives of the protein, said method comprising:
- (i) reacting a polysialic acid at the amine group of the N-terminus of a protein in aqueous acidic solution to produce an N-terminal derivative; and
(ii) purifying the resultant N-terminal derivative in aqueous solution of higher pH than in (i);
wherein the pH of the aqueous solution in (i) is in the range 4.0-6.0 and in step (ii) is in the range 6.5-8.5.
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Abstract
The present invention relates to methods for producing N-terminal derivatives of proteins in which a polysaccharide, preferably having at least terminal sialic units and preferably consisting essentially only of sialic acid units, is reacted at the N-terminus of a protein or peptide under controlled conditions to produce an N-terminal derivative. The controlled conditions include use of acidic pH for the derivatisation step and a higher pH for purification. The derivatives are useful for improving pharmacokinetics and pharmacodynamics of proteins and peptides.
20 Citations
24 Claims
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1. A method for producing a composition comprising a population of polysialic acid derivatives of a protein, wherein the derivatives comprise between 2 and 200 sialic acid units and wherein the population consists substantially only of N-terminal derivatives of the protein, said method comprising:
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(i) reacting a polysialic acid at the amine group of the N-terminus of a protein in aqueous acidic solution to produce an N-terminal derivative; and (ii) purifying the resultant N-terminal derivative in aqueous solution of higher pH than in (i); wherein the pH of the aqueous solution in (i) is in the range 4.0-6.0 and in step (ii) is in the range 6.5-8.5. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 24)
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- 15. A composition comprising a population of polysialic acid derivatives of a protein, wherein the derivatives comprise between 2 and 200 sialic acid units and wherein the population consists substantially only of N-terminal derivatives of the protein.
Specification