US4554157A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Hepatitis B vaccine
Assignee: BTG International Ltd
•
Filing: 1984-05-29
•
Granted: 1985-11-19
•
Expires: 2004-05-29
Abstract
Abstract Immunogenically active fragments of hepatitis B surface antigen, which are a glycoprotein of molecular weight about 28,000 (gp 28) and a protein of molecular weight about 23,000 (p 23), obtained by known methods by treatment of surface antigen with detergent, are prepared in micelle form substantially free from detergent. The micelles are at least as immunogenic as the unfragmented antigen from which they are derived and are useful in the preparation of hepatitis B vaccine. The detergent is removed by layering the detergent containing fragmentation product on an aqueous buffer containing a sucrose gradient and centrifuging the layered buffer when micelles containing both gp 28 and p 23 form and can be recovered in substantially detergent free form from the centrifuged layered buffer.
Inventors
- 👤 Jacinta Skelly
- 👤 Colin R. Howard
- 👤 Arie J. Zuckerman
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-02 11:28
CPC Classifications
A; A61; A61K; A61K39/00; A61K39/12; A61K39/29; A61K39/292; A61K2039/555; A61K2039/55511; A61K2039/55555; C; C12; C12N; C12N2730/00; C12N2730/00011; C12N2730/10011; C12N2730/10111; C12N2730/10134; Y; Y10; Y10S; Y10S530/00; Y10S530/806; Y10S530/808; Y10S530/809; Y10S530/82; Y10S530/826
Claims Analysis
Total Claims: 12
Claim 1
Independent Claim
156 words
In a process for the production of a protein fraction, suitable for use in the formulation of a vaccine against hepatitis B virus infection, which comprises treating serum originating particles of diameter 20-25 nm bearing hepatitis B surface antigen with a non-ionic detergent followed by affinity chromatography to produce a preparation comprising an immunogenic glycopolypeptide of molecular weight about 28,000 (gp 28) and an immunogenic polypeptide of molecular weight about 23,000 (p 23), the improvement which comprises introducing the preparation comprising gp 28 and p 23 and detergent to form a layer on top of an aqueous solution buffered to a pH which avoids denaturation of gp 28 and p 23, said aqueous buffer containing sucrose in a concentration gradient of at least 20% to not more than 65% weight/volume, centrifuging the layered buffer and recovering from the buffer an immunogenic aqueous fraction substantially free from detergent and containing micelles of gp 28 and p 23.
Claim 2
Dependent Claim
156 words
In a process for the production of a protein fraction, suitable for use in the formulation of a vaccine against hepatitis B virus infection, which comprises treating serum originating particles of diameter 20-25 nm bearing hepatitis B surface antigen with a non-ionic detergent followed by affinity chromatography to produce a preparation comprising an immunogenic glycopolypeptide of molecular weight about 28,000 (gp 28) and an immunogenic polypeptide of molecular weight about 23,000 (p 23), the improvement which comprises introducing the preparation comprising gp 28 and p 23 and detergent to form a layer on top of an aqueous solution buffered to a pH which avoids denaturation of gp 28 and p 23, said aqueous buffer containing sucrose in a concentration gradient of at least 20% to not more than 65% weight/volume, centrifuging the layered buffer and recovering from the buffer an immunogenic aqueous fraction substantially free from detergent and containing micelles of gp 28 and p 23.
Claim 3
Dependent Claim
23 words
A process according to claim 1, wherein the concentration gradient of the sucrose is at least 20% but not more than 50% weight/volume.
Claim 4
Dependent Claim
30 words
A process according to claim 1, wherein the concentration gradient of the sucrose in a linear gradient increasing from 20% up to 50% or from 20% up to 60% weight/volume.
Claim 5
Dependent Claim
20 words
A process according to claim 1, 2 or 3, wherein the sucrose containing aqueous buffer has a pH of 7.0-7.5.
Claim 6
Dependent Claim
25 words
A process according to claim 1, 2 or 3, wherein the sucrose containing aqueous buffer includes sodium phosphate, sodium chloride and ethylene diamine tetra-acetic acid.
Claim 7
Dependent Claim
43 words
A process according to claim 1, 2 or 3, wherein the detergent containing polypeptide mixture is introduced as a layer on top of the sucrose containing aqueous buffer and the layered buffer centrifuged at a speed corresponding to a force of 40-250,000 g.
Claim 8
Dependent Claim
18 words
A process according to claim 6, wherein the speed corresponds to a force of at least 200,000 g.
Claim 9
Dependent Claim
20 words
A process according to claim 7, wherein the centrifugation is carried out for at least 6 hours at 4°-20° C.
Claim 10
Dependent Claim
16 words
A process according to claim 8, wherein the centrifugation is carried out for about 24 hours.
Claim 11
Dependent Claim
37 words
A process according to any one of claim 1, 2 or 3, wherein detergent free micelles of gp 28 and p 23 are recovered from the centrifuged layered buffer and formulated in a pyrogen free aqueous medium.
Claim 12
Dependent Claim
14 words
A process according to claim 1, wherein the detergent is an alkaryl polyether alcohol.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1988-11-18 | 1989-05-20 | 1989-11-19 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1992-11-18 | 1993-05-20 | 1993-11-19 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1996-11-18 | 1997-05-20 | 1997-11-19 | $7700.00 | Window Closed / Lapsed |