Patent EP0073656A2 successfully refreshed from Google Patents.
EP0073656A2
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EP
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Preparation of polypeptides in vertebrate cell culture
Assignee: Genentech Inc
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Filing: 1982-08-26
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Granted: —
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Expires: 2002-08-26
Abstract
Abstract A vertebrate cell culture is enabled to produce a desired polypeptide by transformation with a vector whose DNA comprises (a) means for replication in the host; (b) a promoter; and (c) a gene encoding the polypeptide, under the control (b). E.g. a recombinant genome was constructed comprising DNA from (a) the SV40 virus with excision of the VP-1 gene (-hence it is noninfective); (b) the HBsAg gene; and (c) pBR322 (rendering it TET R , and allowing cloning in E. coli). Cells of the CV-1 line of monkey kidney fibroblasts were co-infected therewith and incubated. Lysis occurred, liberating HBsAg which is required for hepatitis vaccine. Non-lytic vectors for expressing HBsAg were also prepared.
Inventors
- 👤 Arthur D. Levinson
- 👤 Daniel G. Yansura
- 👤 Chung-Cheng Liu
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-29 08:50
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/005; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/79; C12N15/85; C12N15/86; A; A61; A61K; A61K39/00; C12N2730/00; C12N2730/00011; C12N2730/10011; C12N2730/10111; C12N2730/10122
Claims Analysis
Total Claims: 18
Claim 1
Independent Claim
133 words
A method of producing a transformant vertebrate cell culture which transformant is capable of producing polypeptide by expression of a replicable expression vector disposed therein encoding said polypeptide, which comprises: a) providing a DNA transfer vector fragment capable of replication in host cell culture, b) providing a DNA fragment comprising a promoter compatible with host cell culture, c) providing a DNA fragment enclosing polypeptide suitable for linkage with the fragment of step b), d) assembling the fragments of steps a), b) and c) together with appropriately positioned transcription ;termination sites and poly A addition sequence and translational start and stop signals to form a replicable expression vector in which said sequence of step c) is under control of said promoter, and e) transforming a vertebrate cell culture with the vector of step d).
Claim 2
Independent Claim
133 words
A method of producing a transformant vertebrate cell culture which transformant is capable of producing polypeptide by expression of a replicable expression vector disposed therein encoding said polypeptide, which comprises: a) providing a DNA transfer vector fragment capable of replication in host cell culture, b) providing a DNA fragment comprising a promoter compatible with host cell culture, c) providing a DNA fragment enclosing polypeptide suitable for linkage with the fragment of step b), d) assembling the fragments of steps a), b) and c) together with appropriately positioned transcription ;termination sites and poly A addition sequence and translational start and stop signals to form a replicable expression vector in which said sequence of step c) is under control of said promoter, and e) transforming a vertebrate cell culture with the vector of step d).
Claim 3
Dependent Claim
36 words
The method according to Claim 1 comprising the additional steps of: f) allowing the vertebrate cell culture transformant to grow under customary conditions in medium until said polypeptide is produced therein, and g) recovering said polypeptide.
Claim 4
Dependent Claim
22 words
The method according to Claim 2 wherein the polypeptide is hepatitis B surface antigen, secreted in discrete particle form into said medium.
Claim 5
Dependent Claim
24 words
The method according to Claim 1, 2 or 3 wherein the fragment of step (a) is further capable of phenotypic selection in microbial host.
Claim 6
Independent Claim
26 words
The method according to any preceding claim whereintbe DNA vector of step a) comprises at.least part of the SV40 vector containing an origin of replication therefor.
Claim 7
Independent Claim
28 words
The method according to any preceding claim wherein the vertebrate cell culture of step e) is either the CV-1 line or the COS-7 line of monkey kidney fibroblasts.
Claim 8
Dependent Claim
63 words
A transformant vertebrate cell culture capable of producing polypeptide by expression of a replicable expression vector disposed therein encoding said polypeptide which includes a replicable DNA expression vector comprising a DNA sequence encoding said polypeptide and an expression promoter compatible with said transformant cell, said sequence being positioned together with translational start and stop signals in said vector under control of said promoter.
Claim 9
Dependent Claim
63 words
A transformant vertebrate cell culture capable of producing polypeptide by expression of a replicable expression vector disposed therein encoding said polypeptide which includes a replicable DNA expression vector comprising a DNA sequence encoding said polypeptide and an expression promoter compatible with said transformant cell, said sequence being positioned together with translational start and stop signals in said vector under control of said promoter.
Claim 10
Dependent Claim
21 words
The cell culture according to Claim 7 obtained by transforming the CV-1 line or the COS-7 line of monkey kidney fibroblasts.
Claim 11
Dependent Claim
36 words
The use of hepatitis B surface antigen, produced by the method according to any of Claims 1 to 6, for conferring immunogenicity to hepatitis B virus in a susceptible human or for preparing a vaccine therefor.
Claim 12
Dependent Claim
24 words
A DNA expression vector capable, in a transformant vertebrate cell culture, of expressing polypeptide under proper reading frame control of an expression effecting promoter.
Claim 13
Dependent Claim
24 words
A DNA expression vector capable, in a transformant vertebrate cell culture, of expressing polypeptide under proper reading frame control of an expression effecting promoter.
Claim 14
Dependent Claim
12 words
A vector according to Claim 10 selected from pSVHBsA, pHBs348-E and pHBs348-L.
Claim 15
Dependent Claim
15 words
A vector according to Claim 10 or 11 capable of expressing hepatitis B surface antigen.
Claim 16
Dependent Claim
25 words
A particle of about 22 nm comprising hepatitis B surface antigen produced in vertebrate cell culture by expression of a replicable expression vector disposed therein,
Claim 17
Dependent Claim
25 words
A particle of about 22 nm comprising hepatitis B surface antigen produced in vertebrate cell culture by expression of a replicable expression vector disposed therein,
Claim 18
Dependent Claim
16 words
The particle according to Claim 13 secreted from said culture into the medium for said culture.
References Cited (Backward Citations)
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