Method for manufacturing non-enveloped virus

Patent metadata

Field	Value
Publication number	US-10072250-B2
Application number	US-201414900837-A
Country	US
Kind code	B2
Filing date	Jul 10, 2014
Priority date	Jul 11, 2013
Publication date	Sep 11, 2018
Grant date	Sep 11, 2018

Abstract

The present invention provides a method for efficiently manufacturing a non-enveloped virus with high purity without laborious operation by cultivating cells having the ability to produce a non-enveloped virus and bringing the cells and an acidic solution into contact with each other. A non-enveloped virus vector manufactured by the method of the present invention and a composition having the non-enveloped viral vector as an active ingredient are very useful as gene transfer methods in the fields of basic research and clinical application gene therapy.

First claim

The invention claimed is:

1. A method of producing a non-enveloped virus, the method comprising:
(a) culturing an isolated cell, wherein a non-enveloped virus is replicating within said cell,
(b) removing the culture solution or media from the cell,
(c) bringing the cell into contact with an acidic solution, and
(d) isolating the non-enveloped virus from the cell and acidic solution mixture of step (c),
wherein the non-enveloped virus is an adeno-associated virus, and wherein the acidic solution is at pH 3.0-6.9.

2. The method according to claim 1 , wherein the acidic solution contains a sodium ion and/or a potassium ion.

3. The method according to claim 1 , wherein the virus is further purified after isolation in step (d).

4. The method according to claim 3 , wherein the purification of the non-enveloped virus is performed by an operation selected from ultracentrifugation, chromatography, and ultrafiltration.

5. The method according to claim 1 , wherein the acidic solution contains a cation and citric acid.

6. The method according to claim 1 , wherein the isolation of step (d) is performed via centrifugation or filtration.

7. The method according to claim 1 , which does not comprise a physical cell disruption method.

Assignees

• Takara Bio Inc

Inventors

• Sakamoto Shuhei
• Kawano Yasuhiro
• Dodo Katsuyuki
• Enoki Tatsuji
• Yoshioka Hirofumi
• Takakura Hikaru
• Mineno Junichi

Classifications

• C12N7/00 Primary

  Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00) · CPC title

• C12N2750/14151

  Methods of production or purification of viral material · CPC title

• C12N2800/50

  Vectors for producing vectors · CPC title

• C12N2750/14351

  Methods of production or purification of viral material · CPC title

• C12N2750/14141

  Use of virus, viral particle or viral elements as a vector · CPC title