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AssigneeNovo Nordisk A/S
InventorsHenrik Dalb.o slashed.ge, John Pedersen, Thorkild Christensen, J.o slashed.rli Winnie Ringsted, Torben Ehlern Jessen
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A desired protein having the formula: A-B-C-P wherein a) A is Lys or Arg, and B and C are arbitrary amino acids, or b) A is an arbitrary amino acid different from Pro, Lys and Arg, and B and/or C is Pro, is produced from a biosynthetically formed amino acid extended protein having the formula: X-A-B-C-P wherein A, B, C and P are as defined above, and X is an amino acid sequence with an even number of amino acids, of which the first one, seen from the N-terminal end, is different from Lys and Arg, all other uneven amino acids are different from Pro, Lys and Arg, and all even amino acids are different from Pro, by reaction with the enzyme dipeptidyl aminopeptidase (DAP I). The desired protein is obtained in a pure state. Thus, e.g. hGH without content of Met-hGH may be produced by the process.
A process for preparing ripe proteins, which comprises expressing in a procaryotic or eukaryotic cell a DNA sequence which codes for the synthesis of a fusion protein which comprises the amino acid sequence of the desired ripe protein and a N-terminal amino acid pro-sequence, cleaving the pro-sequence using DAP I and recovering the desired ripe protein.
A process for separating a desired ripe protein from a fusion protein comprising the desired ripe protein having a negatively charged amino acid sequence of suitable length fused to the N-terminus of said desired ripe protein, said amino acid sequence comprising an even number of amino acids, which process comprises cleaving the amino acid sequence using dipeptidyl aminopeptidase I under suitable cleavage conditions, and recovering the desired ripe protein utilizing the existence of the negative charge in the amino acid sequence.
A process for separating a desired ripe protein from a fusion protein comprising the desired ripe protein having a charged amino acid sequence of suitable length fused to the N- terminus of said desired ripe protein wherein at least one of the charged amino acids is attached to the N-terminal of said desired ripe protein, said amino acid sequence comprising an even number of amino acids, which process comprises cleaving the amino acid sequence using dipeptidyl aminopeptidase I under suitable cleavage conditions, and recovering the desired ripe protein utilizing the existence of the charge in the charged amino acid sequence.
A process for preparing a desired ripe protein which comprises expressing in a procaryotic or eukaryotic cell a DNA sequence which codes for the synthesis of a fusion protein which comprises the amino acid sequence of the desired ripe protein and an N-terminal amino acid pro-sequence, cleaving the pro-sequence using an dipeptidyl aminopeptidase and recovering the desired ripe protein.
A method of producing a protein consisting of amine acids 1-191 of human growth hormone comprising:
A process for producing ripe human growth hormone comprising
Dependencies are inferred from the claim text.