US2004115775A1PendingUtilityA1
Homogeneity and secretion of recombinant proteins in mammalian systems
Priority: Dec 5, 2000Filed: Dec 5, 2001Published: Jun 17, 2004
Est. expiryDec 5, 2020(expired)· nominal 20-yr term from priority
C07K 2319/75C12N 15/625C07K 2319/02
37
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Claims
Abstract
The present invention relates to a method for improving homogeneity and/or secretion of a recombinant protein of interest expressed in mammalian cells by replacing the endogenous signal peptide sequence of the DNA encoding the protein of interest with that of human hGH. The invention also relates to DNA expression vectors containing the sequence encoding such proteins fused to the signal peptide sequence of the hGH and to cells harboring such vectors.
Claims
exact text as granted — not AI-modified1 . A method for improving homogeneity and/or secretion of a recombinant protein of interest expressed in a mammalian system comprising replacing the endogenous signal peptide sequence of the DNA encoding the protein of interest with that of hGH.
2 . A method according to claim 1 , wherein the protein of interst IL-18BP.
3 . A method according to claim 2 , wherein the mammalian system comprises CHO cells.
4 . A method according to claim 3 , wherein the CHO cells are cultured in medium supplemented with serum.
5 . A method according to claim 3 , wherein the CHO cells are cultured under serum free conditions.
6 . An expression vector for improving homogeneity and/or secretion of a recombinant protein of interest expressed in a mammalian system comprising the signal peptide sequence of hGH and the DNA encoding the protein of interest.
7 . An expression vector according to claim 6 , which directs expression of a homogenous protein.
8 . An expression vector according to claim 6 , which directs efficient expression of the protein of interest.
9 . An expression vector according to claim 6 , which directs efficient translation of the protein of interest.
10 . An expression vector according to claim 6 , in which the signal peptide encodes an optimal Kozalk sequence.
11 . An expression vector according to claim 6 , which directs efficient secretion of the protein of interest.
12 . An expression vector according to claim 6 , which allows constitutive expression of the protein of interest.
13 . An expression vector according to claim 6 , wherein the expression of the gene encoding the protein of interest is regulated by the CMV promoter.
14 . An expression vector according to claim 6 , wherein the expression of the gene encoding the protein of interest is regulated by the polyadenylation signal of the human growth hormone.
15 . An expression vector in accordance with anyone of claims 6 to 14 , wherein the protein of interest is IL-18BP.
16 . A cell harbouring a vector according to anyone of claims 6 to 15 .
17 . A cell according to claim 16 , producing about 12 picogram IL-18BP/cell/24 hours.
18 . A cell according to claim 16 , producing 11.8 picogram IL-18BP/cell/24 hours.
19 . A cell according to claim 16 , producing at least 4 picogram IL-18BP/cell/24 hours.
20 . A cell according to claim 16 , producing at least 11.8 picogram IL-18BP/cell/24 hours.
21 . A cell according to claim 16 , capable of growing in serum free medium.
22 . A cell according to claim 21 , producing at least 4 picogram IL-18 BP/cell/24 hours.
23 . A cell according to claim 21 , producing about 12 picogram IL-18 BP/cell/24 hours.
24 . A cell according to claim 21 , producing 11.8 picogram IL-18BP/cell/24 hours.
25 . A cell according to claim 21 , producing at least 11.8 picogram IL-18BP/cell/24 hours.
26 . A protein obtainable by the method of claim 1 .
27 . A protein according to claim 26 , being IL-18BP.
28 . A pharmaceutical composition comprising a protein according to claims 26 or 27 .Join the waitlist — get patent alerts
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