Patent US4522809A successfully refreshed from Google Patents.
US4522809A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Process for obtaining lipid envelope virus sub-units, notably antigens for use as vaccines, the products obtained and their applications
Assignee: Institut Pasteur
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Filing: 1984-08-01
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Granted: 1985-06-11
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Expires: 2004-08-01
Abstract
Abstract A process is disclosed for producing virus sub-units with lipid envelopes, comprising, in a neutral or basic pH, dissolving a lower halogenated hydrocarbon, at a concentration equivalent to or approximating its limit of solubility, in an aqueous virus suspension, thereafter contacting, with stirring, the preparation so obtained with a nonionic detergent at a minimum concentration effective to induce disruption of virions into heavy sub-units for a length of time sufficient to permit said disruption, and in separating the heavy sub-units so obtained from the reaction medium. The process permits preparation of purified disrupted antigens for use as vaccines, notably influenza vaccines.
Inventors
- 👤 Philippe Adamowicz
- 👤 Ludwig Muller
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-07 11:45
CPC Classifications
C; C12; C12N; C12N7/00; A; A61; A61K; A61K39/00; A61K39/12; A61K39/145; A61K2039/51; A61K2039/525; A61K2039/5252; A61K2039/70; C12N2760/00; C12N2760/00011; C12N2760/16011; C12N2760/16111; C12N2760/16134; C12N2760/16161; C12N2760/16163; C12N2760/16211; C12N2760/16234
Claims Analysis
Total Claims: 10
Claim 1
Independent Claim
192 words
A process for the production of immunogenic heavy subunits of influenza virus, comprising the following steps, carried out at a neutral or basic pH: (1) dissolving an amount of chloroform in an aqueous influenza virus suspension to obtain a concentration of chloroform equivalent to or approximating its solubility limit in said suspension, thereafter, (2) adding, with stirring, to the preparation so obtained a nonionic detergent while maintaining a single phase system, said detergent being added at the minimal concentration necessary to effect disruption of the virus into heavy immunogenic subunits, and stirring this mixture for a sufficient length of time to obtain said disruption, said heavy subunits comprising neuraminidase and hemagglutinin antigens in their original arrangement attached to fragments of the lipid envelopes of said virus, and (3) separating the heavy subunits so obtained from lighter components by applying gradient fractionation methods to the reaction mixture, said lighter components comprising said chloroform, said detergent and other products originating from the disruption of said virus, and said heavy subunits being in the dense fraction of the gradient, wherein an inactivation agent is added to the reaction medium at any time during the process.
Claim 2
Dependent Claim
192 words
A process for the production of immunogenic heavy subunits of influenza virus, comprising the following steps, carried out at a neutral or basic pH: (1) dissolving an amount of chloroform in an aqueous influenza virus suspension to obtain a concentration of chloroform equivalent to or approximating its solubility limit in said suspension, thereafter, (2) adding, with stirring, to the preparation so obtained a nonionic detergent while maintaining a single phase system, said detergent being added at the minimal concentration necessary to effect disruption of the virus into heavy immunogenic subunits, and stirring this mixture for a sufficient length of time to obtain said disruption, said heavy subunits comprising neuraminidase and hemagglutinin antigens in their original arrangement attached to fragments of the lipid envelopes of said virus, and (3) separating the heavy subunits so obtained from lighter components by applying gradient fractionation methods to the reaction mixture, said lighter components comprising said chloroform, said detergent and other products originating from the disruption of said virus, and said heavy subunits being in the dense fraction of the gradient, wherein an inactivation agent is added to the reaction medium at any time during the process.
Claim 3
Dependent Claim
71 words
Process according to claim 1, which comprises the following steps: (1) dissolving said chloroform to obtain a concentration of about 0.4 to 0.7% (volume/volume), (2) adding said nonionic detergent to obtain a concentration in the range of about 0.02 to 0.2% and stirring the reaction medium, at a pH between about 7.5 and about 8.5 for 1 to 20 hours, and (3) separating the immunogenic heavy subunits of the influenza virus.
Claim 4
Dependent Claim
112 words
Process according to claim 1 for obtaining a preparation of purified disrupted antigens of influenza virus, which comprises the following steps of: (1) adding said chloroform at a concentration of between about 0.4 and 0.5% (volume/volume) to influenza virus suspension; (2) adding, when the chloroform is completely dissolved and after adjusting the pH to a value of between 8 and 8.7, said nonionic detergent at a concentration of about 0.02 to 0.2%, (3) adding an additional amount of chloroform of up to 0.2%, (4) stirring the resulting reaction mixture, which is in the form of a monophasic system, for 1 to 20 hours, and (5) in purifying the preparation of split virions.
Claim 5
Dependent Claim
28 words
Process according to claim 1, wherein the inactivation agent is β-propiolactone and it is added after the addition of the nonionic detergent at a rate of about 0.02%.
Claim 6
Dependent Claim
60 words
Process according to claim 1, wherein said aqueous suspension of influenza virus is obtained from an allantonic solution of influenza virus which, after its preparation, is subjected in a known manner to adsorption-elution steps on erythrocytes, then clarification by centrifugation and filtration, thus providing a concentrate containing the virion, which is used to form the aqueous solution as starting material.
Claim 7
Dependent Claim
27 words
Process according to claim 1 wherein said nonionic detergent is a surfactant containing polyoxyethylenic derivatives of partial esters of fatty acids and hexitol anhydrides derived from sorbitol.
Claim 8
Dependent Claim
56 words
Process according to claim 1, wherein said nonionic detergent is selected from the class consisting of: (1) condensates of alkylphenols and ethylene oxide; (2) addition products of aliphatic ethers and ethylene oxide; (3) addition products of esters and ethylene oxide; (4) addition products of amines and ethylene oxide; (5) addition products of alkanolamides and ethylene oxides.
Claim 9
Dependent Claim
37 words
A process according to claim 1, wherein an additional amount of chloroform up to the amount of step (1) is added after said nonionic detergent to the reaction medium at a pH between about 8 and 8.7.
Claim 10
Dependent Claim
21 words
A process according to claim 8, wherein said chloroform is in an amount up to 0.2% (volume/volume) of the reaction medium.
References Cited (Backward Citations)
FR1587316A
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US3847737A
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FR2248054A1
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FR2422720A2
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US4199450A
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FR2475572A1
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