US4803164A
Expired / Lapsed
US
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Preparation of hepatitis b surface antigen in yeast
Assignee: Genentech Inc
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Filing: 1987-04-22
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Granted: 1989-02-07
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Expires: 2007-04-22
Abstract
Abstract Hepatitis surface antigen is synthesized in recombinant yeast hosts transformed with vectors encoding hepatitis surface antigen, preferably under the control of the yeast PGK promoter and preferably in the absence of DNA encoding the surface antigen precursor. Hepatitis surface antigen is assembled by yeast into antigenic 22 nm particles even though hepatitis surface antigen bacterial transformants were not known to be capable of assembling the surface antigen into 22 nm particles.
Inventors
- 👤 Ronald A. Hitzeman
- 👤 Arthur D. Levinson
- 👤 Daniel G. Yansura
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-09 23:10
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/005; A; A61; A61K; A61K39/00; A61K39/12; A61K39/29; A61K39/292; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/79; C12N15/80; C12N15/81; C12N2730/00; C12N2730/00011; C12N2730/10011; C12N2730/10111; C12N2730/10122; C12N2730/10134
Claims Analysis
Total Claims: 13
Claim 1
Independent Claim
88 words
A DNA expression vector capable of replication and phenotypic selection in yeast host strain comprising a promoter compatible with a yeast host strain and a DNA sequence encoding hepatitis B surface antigen, said sequence being positioned together with translational start and stop signals in said vector under control of said promoter such that in a transformant yeast strain it is expressed to produce hepatitis B surface antigen in particle form having a sedimentation rate which is virtually identical to that of authentic 22 nm hepatitis surface antigen particles.
Claim 2
Dependent Claim
88 words
A DNA expression vector capable of replication and phenotypic selection in yeast host strain comprising a promoter compatible with a yeast host strain and a DNA sequence encoding hepatitis B surface antigen, said sequence being positioned together with translational start and stop signals in said vector under control of said promoter such that in a transformant yeast strain it is expressed to produce hepatitis B surface antigen in particle form having a sedimentation rate which is virtually identical to that of authentic 22 nm hepatitis surface antigen particles.
Claim 3
Dependent Claim
13 words
A yeast strain transformed with the DNA expression vector according to claim 1.
Claim 4
Dependent Claim
15 words
The yeast strain according to claim 2 obtained by transforming a leu2 auxotrophic yeast strain.
Claim 5
Dependent Claim
12 words
The yeast strain according to claim 2 obtained by transforming strain XV610-8C.
Claim 6
Dependent Claim
14 words
A fermentation culture comprising a transformed yeast according to claim 2, 3, or 4.
Claim 7
Dependent Claim
24 words
The vector of claim 1 wherein the DNA sequence encoding hepatitis B surface antigen encodes only the mature hepatitis B surface antigen structural gene.
Claim 8
Independent Claim
168 words
A method of producing hepatitis B surface antigen in particle form suitable for use in conferring immunogenicity to hepatitis B virus in a susceptible human which comprises: (a) providing a DNA transfer vector capable of replication and phenotypic selection in yeast host strains, (b) providing a DNA fragment comprising a promoter compatible with a yeast host strain, (c) providing a DNA fragment encoding hepatitis B surface antigen, (d) assembling the fragments of steps (a), (b) and (c) together with translational start and stop signals for the fragment of step (c) to form a replicable expression vector so that said sequence of step (c) is under control of said promoter, (e) transforming a yeast strain with the vector of step (d), (f) allowing the yeast transformant to grow under fermentation conditions until said hepatitis B surface antigen is produced therein, and (g) recovering said hepatitis B surface antigen in particle form having a sedimentation rate which is virtually identical to that of authentic 22 nm hepatitis surface antigen particles.
Claim 9
Independent Claim
168 words
A method of producing hepatitis B surface antigen in particle form suitable for use in conferring immunogenicity to hepatitis B virus in a susceptible human which comprises: (a) providing a DNA transfer vector capable of replication and phenotypic selection in yeast host strains, (b) providing a DNA fragment comprising a promoter compatible with a yeast host strain, (c) providing a DNA fragment encoding hepatitis B surface antigen, (d) assembling the fragments of steps (a), (b) and (c) together with translational start and stop signals for the fragment of step (c) to form a replicable expression vector so that said sequence of step (c) is under control of said promoter, (e) transforming a yeast strain with the vector of step (d), (f) allowing the yeast transformant to grow under fermentation conditions until said hepatitis B surface antigen is produced therein, and (g) recovering said hepatitis B surface antigen in particle form having a sedimentation rate which is virtually identical to that of authentic 22 nm hepatitis surface antigen particles.
Claim 10
Dependent Claim
48 words
The method according to claim 7 wherein the DNA fragment of step (c) comprises, in order from the 5' end of its coding strand, a ATG translational start codon, the nucleotides encoding hepatitis B surface antigen of the hepatitis B genome, and one or more translational stop signals.
Claim 11
Dependent Claim
15 words
The method according to claim 7 wherein the yeast strain of step (e) is XV610-8C.
Claim 12
Independent Claim
25 words
The method according to any one of claims 7 to 9 wherein the promoter of step (b) is derived from the yeast PGK promoter region.
Claim 13
Dependent Claim
21 words
The method of claim 8 wherein the start codon is the start codon for mature hepatitis B surface antigen structural gene.
References Cited (Backward Citations)
FR2270892A1
Cited
EP0011562A1
Cited
GB2034323A
Cited
EP0013828A1
Cited
EP0020251A2
Cited
GB2068969A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1992-02-07 | 1992-08-08 | 1993-02-07 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1996-02-07 | 1996-08-08 | 1997-02-07 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2000-02-07 | 2000-08-08 | 2001-02-07 | $7700.00 | Window Closed / Lapsed |