US4402939A
Expired / Lapsed
US
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Vaccinating glycopeptidic antigenic fraction with a very high level of immunogenicity, isolated from cultures of pathogenic germs, processes for isolating said fraction and vaccines containing said fraction
Assignee: Institut Pasteur
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Filing: 1980-06-26
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Granted: 1983-09-06
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Expires: 2000-06-26
Abstract
Abstract A glycopeptic vaccinating antigenic fraction with a very high level of immunogenicity, which is isolated from a culture of pathogenic germs comprises an osidic part present in a proportion of about 50%, with a proteinic part, which are linked together by covalent bond; when it is opposed to a suitable immune-serum, the fraction has an immunological image which essentially comprises a 'clear' line of precipitation close to the antigen pit, in immunodiffusion in gelose. The fraction is used for the preparation of vaccines and as a diagnostic agent.
Inventors
- 👤 Jean-Michel Fournier
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-08 21:31
CPC Classifications
C; C12; C12P; C12P21/00; C12P21/005; A; A61; A61K; A61K39/00; A61K39/02; A61K39/025; A61K39/0266; A61K39/09; A61K39/092; C07; C07K; C07K14/00; C07K14/195; C07K14/24; C07K14/26
Claims Analysis
Total Claims: 22
Claim 1
Independent Claim
84 words
A process for extracting and purifying a vaccinating antigenic fraction, comprising: extracting a bacterial deposit one to several times, with very vigorous agitation, by a suitable buffer with a pH-value of close to 6, said bacteria being a pathogenic germ which neither secretes nor excretes toxins, and which contains a protective antigen defined by a line of precipitation centered on the antigen well, in immunodiffusion in gelose, when opposed to an immune-serum from an animal treated by a culture of the same pathogenic germ;
Claim 2
Dependent Claim
84 words
A process for extracting and purifying a vaccinating antigenic fraction, comprising: extracting a bacterial deposit one to several times, with very vigorous agitation, by a suitable buffer with a pH-value of close to 6, said bacteria being a pathogenic germ which neither secretes nor excretes toxins, and which contains a protective antigen defined by a line of precipitation centered on the antigen well, in immunodiffusion in gelose, when opposed to an immune-serum from an animal treated by a culture of the same pathogenic germ;
Claim 3
Dependent Claim
48 words
concentrating the supernatant obtained containing the crude surface constituents by ultra-filtration, collecting only the particles whose molecular weight is more than 10,000; treating the ultrafiltrate produced with sodium deoxycholate (DOC) in a suitable buffer with a pH-value of close to 8; precipitating the treated ultra-filtrate with absolute ethanol;
Claim 4
Dependent Claim
48 words
concentrating the supernatant obtained containing the crude surface constituents by ultra-filtration, collecting only the particles whose molecular weight is more than 10,000; treating the ultrafiltrate produced with sodium deoxycholate (DOC) in a suitable buffer with a pH-value of close to 8; precipitating the treated ultra-filtrate with absolute ethanol;
Claim 5
Dependent Claim
54 words
dissolving the precipitate produced with the same buffer with a pH-value of close to 8 with the addition of 2% of DOC; dializing the obtained solution against the buffer Tris-HCl with a pH-value of 8; adsorbing on DEAE-cellulose; eluting the antigen with the same buffer used with increasing molarity; and then concentrating by ultra-filtration.
Claim 6
Dependent Claim
54 words
dissolving the precipitate produced with the same buffer with a pH-value of close to 8 with the addition of 2% of DOC; dializing the obtained solution against the buffer Tris-HCl with a pH-value of 8; adsorbing on DEAE-cellulose; eluting the antigen with the same buffer used with increasing molarity; and then concentrating by ultra-filtration.
Claim 7
Dependent Claim
49 words
A process according to claim 1, wherein said extracting step comprises extracting with a buffer comprising: ______________________________________
lithium acetate 0.2 M
EDTA 10 mM
sodium azide 0.02%
acetic acid QSP
→ pH-value 6
______________________________________ in proportion of 50 to 70 ml of buffer per gram of wet bacterial deposit.
Claim 8
Dependent Claim
29 words
A process according to claim 1, further including the step of filtering the solution of crude surface constituents on a Millipore filter before said ultra-filtration step is carried out.
Claim 9
Dependent Claim
71 words
A process according to claim 1, wherein said treating step comprises treating the ultra-filtrate, which is adjusted to a pH-value of close to 8 by the buffer Tris, with a 5% solution of DOC in buffer Tris-HCl with a pH-value of 8, formed by: ______________________________________
Tris-HCl 10 mM
EDTA 10 mM
______________________________________ so as to adjust the concentration of the solution of the surface constituents to about 1% (weight/volume) of DOC.
Claim 10
Dependent Claim
25 words
A process according to claim 1, wherein said precipitating step comprises precipitating the treated ultra-filtrate by four to five times its volume of absolute alcohol.
Claim 11
Dependent Claim
35 words
A process according to claim 1, further including the step of calibrating the tubes containing the antigen after passing over a DEAE-cellulose column by the protection test and/or the immunodiffusion test and/or the immunoelectrophoresis test.
Claim 12
Dependent Claim
30 words
A process for extracting and purifying the vaccinating antigenic fraction according to claim 1 comprising producing said fraction by purification of a preparation containing said surface antigens, by affinity chromatography.
Claim 13
Dependent Claim
40 words
A process according to claim 7, wherein said purification by affinity chromatography is performed using an immunoadsorbent formed by antibodies which are specific of the component which forms the `clear` line precipitate, which are rendered insoluble by any suitable means.
Claim 14
Dependent Claim
40 words
A process according to claim 8, wherein said specific antibodies are produced by hyperimmunization of mammals such as the rabbit in particular, by a preparation of surface antigens of pathogenic germs, in respect of which immunization is to be obtained.
Claim 15
Dependent Claim
34 words
A process according to claim 9, wherein said specific antibodies are produced by injection into an animal to be immunized of preparations of surface antigens which are purified by per se known biochemical methods.
Claim 16
Dependent Claim
80 words
A process according to claim 9 wherein said specific antibodies are produced by the direct use of the `clear` line precipitate which is formed in the gelose in the course of the immunodiffusion reaction, which precipitate is recovered by cutting out in situ in the gelose, advantageously after washing of said gelose to remove the antigens or the antibodies which are not complexed in the course of the immunodiffusion reaction, whereafter it is injected in the animal to be immunized.
Claim 17
Dependent Claim
20 words
A process according to claim 11, wherein the clear line precipitate is produced using purified specific reagents of the determinant.
Claim 18
Dependent Claim
24 words
A process according to claim 11, wherein the clear line precipitate is produced using preparations of antigens and antibodies, which have not been purified.
Claim 19
Dependent Claim
52 words
A process according to claim 8, wherein the specific antibodies used as immunoadsorbents, in the method for purification of the antigens by affinity chromatography, are recovered after having collected the blood of the hyperimmunised animals and after having separated from the red blood corpuscles, the serum which is rich in specific antibodies.
Claim 20
Dependent Claim
18 words
A process according to claim 8, wherein the antibodies are rendered insoluble by polymerisation by means of glutaraldehyde.
Claim 21
Dependent Claim
27 words
A process according to claim 8, wherein the antibodies are rendered insoluble by fixing on beads of suitable polymers or on glass beads which are suitably activated.
Claim 22
Dependent Claim
76 words
A process according to claim 7, wherein the preparation of surface antigens which is subjected to purification by affinity chromatography is selected from the group which comprises bacteria cultures or the supernatants of such cultures, or the supernatants containing the crude surface constituent, which are produced by extraction of a bacterial deposit, or by the ultra-filtrate produced after ultra-filtration of the supernatant containing the crude surface constituent, and treatment of the latter by sodium deoxycholate (DOC).
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1986-09-05 | 1987-03-07 | 1987-09-06 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1990-09-05 | 1991-03-07 | 1991-09-06 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1994-09-05 | 1995-03-07 | 1995-09-06 | $7700.00 | Window Closed / Lapsed |