Patent EP0161937A2 successfully refreshed from Google Patents.
EP0161937A2
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EP
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Recombinant fusion proteins
Assignee: Celltech R&D Ltd
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Filing: 1985-05-15
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Granted: —
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Expires: 2005-05-15
Abstract
Abstract This invention provides a DNA sequence coding for a cleavage site which is specifically cleaved by blood coagulation Factor Xa, a vector containing such a sequence, and a host organism transformed with such a vector. Preferably, in the vector, the Factor Xa cleavage site coding sequence is fused at one end to a product and at its other end to an ATG codon or a sequence coding for at least part of a host protein. This invention also provides a process, for the production of a desired protein or peptide product in native form, comprising: transforming a host organism with a vector as described above; expressing the desired protein or peptide product as a fusion protein comprising the desired protein or peptide product fused to a Factor Xa cleavage site; and cleaving the fusion protein with Factor Xa to yield the foreign gene product in native form.
Inventors
- 👤 Kiyoshi Nagai
- 👤 Hans Christian Thogersen
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-26 22:44
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/10; C12N9/1025; C12N9/1029; C12N9/1033; C07; C07K; C07K14/00; C07K14/435; C07K14/46; C07K14/47; C07K14/4701; C07K14/4716; C07K14/575; C07K14/585; C07K14/795; C07K14/805; C12N15/00; C12N15/09; C12N15/11; C12N15/62; A; A61; A61K; A61K38/00; C07K2319/00; C07K2319/50; C07K2319/61; C07K2319/70; C07K2319/74; C07K2319/75
Claims Analysis
Total Claims: 25
Claim 3
Dependent Claim
17 words
A DNA sequence coding for a cleavage site which is specifically cleaved by blood coagulation Factor Xa.
Claim 4
Dependent Claim
17 words
A DNA sequence coding for a cleavage site which is specifically cleaved by blood coagulation Factor Xa.
Claim 5
Dependent Claim
28 words
The DNA sequence of claim 1, which codes for the amino acid sequence X-Y-Gly-Arg wherein X is Ile,Leu,Pro or Ala, and Y is Glu, Asp, Gln or Asn.
Claim 6
Dependent Claim
16 words
The DNA sequence of claim 2, wherein X is Ile and Y is Glu or Gln.
Claim 7
Dependent Claim
14 words
The DNA sequence of claim 3, which codes for the amino acid sequence Ile-Glu-Gly-Arg.
Claim 8
Dependent Claim
13 words
The DNA sequence of claim 4, containing the sequence ATC GAG GGT AGG.
Claim 9
Dependent Claim
14 words
The DNA sequence of claim 5, containing the sequence ATC GAG GGT AGG CCT.
Claim 10
Dependent Claim
13 words
The DNA sequence of claim 4, containing the sequence ATT GAA GGT CGT.
Claim 11
Dependent Claim
16 words
A vector comprising a DNA coding sequence according to any one of claims 1 to 7.
Claim 12
Dependent Claim
10 words
The vectors M13mp11FX, M13mpII FX-MCL pLcIIFXβ, pLcIIFX-MLC, and pCT 20210.
Claim 13
Dependent Claim
10 words
The vectors M13mp11FX, M13mpII FX-MCL pLcIIFXβ, pLcIIFX-MLC, and pCT 20210.
Claim 14
Dependent Claim
37 words
The vector of claim 8, wherein the DNA sequence coding for a cleavage site which is specifically cleaved by blood coagulation Factor Xa is fused to a DNA sequence coding for a desired protein or peptide product.
Claim 15
Dependent Claim
23 words
The vector of claim 10, wherein the DNA sequence coding for the Factor Xa cleavage site is also fused to an ATG codon.
Claim 16
Dependent Claim
31 words
The vector of claim 10, wherein the DNA sequence coding for the Factor Xa cleavage site is also fused to a sequence coding for at least part of a host protein.
Claim 17
Dependent Claim
18 words
The vector of claim 12, wherein the host protein is lacZ, tufB, trpE, trpD, bla, λCII or CAT.
Claim 18
Dependent Claim
39 words
The vector of any one of claims 10 to 13, wherein the DNA sequence coding for the Factor Xa cleavage site is fused to the N-terminal end of the DNA sequence coding for the desired protein or peptide product.
Claim 19
Dependent Claim
30 words
The vector of any one of claims 10 to 14, wherein the desired protein or peptide product is an enzyme, a serum protein, a peptide hormone, or a precursor thereof.
Claim 20
Dependent Claim
19 words
The vector of claim 15, wherein the desired protein or peptide product is P-globin, calcitonin-glycine, or myoglobin light chain.
Claim 21
Dependent Claim
16 words
A host organism transformed with a vector according to any one of claims 8 to 16.
Claim 22
Dependent Claim
18 words
An E.coli or Bacillus cell transformed with a vector according to any one of claims 8 to 16.
Claim 23
Dependent Claim
19 words
MZ-1 or E. coli HB101 cells transformed with a vector according to any one of claims 8 to 16.
Claim 24
Dependent Claim
73 words
A process, for the production of a desired protein or peptide product in native form, comprising: transforming a host organism with a vector according to any one of claims 10 to 16; expressing the desired protein or peptide product as a fusion protein comprising the desired protein or peptide product fused to a Factor Xa cleavage site; and cleaving the fusion protein with Factor Xa to yield the-foreign gene product in native form.
Claim 25
Dependent Claim
15 words
The process of claim 20, wherein the Factor Xa is prepared by activating Factor X
Claim 26
Dependent Claim
21 words
The process of claim 20 or claim 21 wherein the Factor X is bovine Factor X 1 or X 2 .
Claim 27
Dependent Claim
64 words
The process of any one of claims 20 to 22, wherein the fusion protein comprises at least a part of a host protein linked to the Factor Xa cleavage site, the host protein being purifiable by affinity chromatography, and including the step of purifying the fusion protein by use of an affinity chromotography medium specific for the host protein prior to the cleavage step.
References Cited (Backward Citations)
No citation records indexed.
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