# US 10,946,088 B2 · Preparation of influenza virus vaccine antigens

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- Number: US 10,946,088 B2
- Document ID: US10946088B2
- Status: Granted
- Kind: B2
- Application number: 14/961,778
- Filed: 2015-12-07
- Published: 2016-06-09
- Granted: 2021-03-16
- Earliest claimed: 2008-03-18
- Assignees: Seqirus UK Limited
- Applicants: Seqirus UK Limited
- Inventors: Christoph Haussmann; Frank Hauschild; Bjorn Jobst
- Web page: https://patentgrep.com/patent/US10946088B2

## CPC

- A61K 39/145: Orthomyxoviridae, e.g. influenza virus
- A61K 39/12: Viral antigens
- A61P 31/12: Antivirals
- A61P 31/16: for influenza or rhinoviruses
- A61P 37/04: Immunostimulants
- C12N 7/00: Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00)
- A61K 2039/5252: {inactivated (killed)}
- C12N 2760/16034: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
- C12N 2760/16051: Methods of production or purification of viral material
- C12N 2760/16134: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
- C12N 2760/16163: by chemical treatment
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

A number of improvements for preparing vaccine antigens from disintegrated influenza viruses are disclosed. A splitting step can be followed by detergent exchange. Splitting can take place in the presence of a buffer with a higher ionic strength and/or in the presence of phosphate buffer.

## Claims

### Claim 1

A method for disrupting influenza virions, the method comprising steps of:
(i) obtaining a composition comprising influenza virions prepared from a cell culture,
(ii) inactivating the influenza virions to obtain inactivated virions, and
(iii) splitting the inactivated virions in the presence of a buffer having an ionic strength of from 200 mM to 800 mM, wherein the buffer comprises phosphate.

### Claim 2 (depends on 1)

The method of claim 1, wherein the influenza virions are from an influenza A virus.

### Claim 3 (depends on 1)

The method of claim 1, wherein the influenza virions are from an influenza A virus of H5 hemagglutinin subtype.

### Claim 4 (depends on 1)

The method of claim 1, wherein the cell culture is a mammalian cell culture or an avian cell culture.

### Claim 5 (depends on 1)

A method for manufacturing an influenza vaccine, the method comprising steps of:
carrying out the method of claim 1,
obtaining a bulk antigen preparation from the disrupted virions of step (iii), and
formulating the bulk antigen preparation into an influenza vaccine.

### Claim 6 (depends on 5)

The method of claim 5, wherein the influenza virions are from an influenza A virus.

### Claim 7 (depends on 5)

The method of claim 5, wherein the influenza virions are from an influenza A virus of H5 hemagglutinin subtype.

### Claim 8 (depends on 5)

The method of claim 5, wherein the cell culture is a mammalian cell culture or an avian cell culture.

### Claim 9 (depends on 5)

The method of claim 5, wherein the influenza vaccine contains less than 10 ng host cell DNA.

### Claim 10 (depends on 5)

The method of claim 5, wherein the influenza vaccine is a monovalent or trivalent influenza vaccine.

### Claim 11 (depends on 5)

The method of claim 5, wherein the influenza vaccine comprises hemagglutinin from at least one B strain.

### Claim 12 (depends on 5)

The method of claim 5, wherein the influenza vaccine comprises from about 3.75 μg to about 15 μg hemagglutinin, per strain, per dose.

### Claim 13 (depends on 5)

The method of claim 5, wherein the step of formulating comprises combining the bulk antigen preparation with an adjuvant.

### Claim 14 (depends on 13)

The method of claim 13, wherein the adjuvant is an oil-in-water emulsion.

### Claim 15 (depends on 1)

The method of claim 1, wherein the buffer has an ionic strength of at least 300 mM.

### Claim 16 (depends on 1)

The method of claim 1, wherein the buffer has an ionic strength of at least 400 mM.

### Claim 17 (depends on 1)

The method of claim 1, wherein the buffer has an ionic strength of at least 500 mM.

### Claim 18 (depends on 1)

The method of claim 1, wherein the buffer has a pH of from 6.5 to 8.5.

### Claim 19 (depends on 1)

The method of claim 1, wherein the buffer has a pH of from 7.0 to 8.0.

### Claim 20 (depends on 1)

The method of claim 1, wherein the buffer has a pH of from 7.0 to 7.8.

### Claim 21 (depends on 1)

The method of claim 1, wherein the method results in a higher than 20-fold increase in antigen yield, when compared to a method for disrupting influenza virions comprising corresponding steps (i)-(iii), except that step (iii) is performed in the presence of a Tris buffer.

### Claim 22 (depends on 1)

The method of claim 1, wherein the method results in an increased antigen yield, when compared to a method for disrupting influenza virions comprising corresponding steps (i)-(iii), except that the buffer in step (iii) has an ionic strength of less than 200 mM.