IP Library › Granted Patent US 12,398,375
Granted Patent B2
US 12,398,375 · App. 17/266,499 · Granted Aug 26, 2025

Methods for improved poxvirus yields

Inventors: Douglas Grant McFadden (Tempe, AZ); Mohammed Masmudur Rahman (Chandler, AZ)
Assignee: ARIZONA BOARD OF REGENTS ON BEHALF OF ARIZONA STATE UNIVERSITY
C12N7/00A61P31/00C12N2710/24051C12N2710/24151
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Quick Facts
Patent No.
US 12,398,375
App. No.
17/266,499
Granted
Aug 26, 2025
Kind
B2
Abstract

A method of producing poxviruses at an increased growth rate and/or progeny virus titer in cells, the method including: contacting a host cell with an effective amount of nucleocytoplasmic transport inhibitor; contacting the host cell with a poxvirus of interest under conditions that permit the poxvirus of interest to adsorb to the surface of the host cell; and culturing the host cell to produce progeny poxvirus of interest.

Claims (23)

1. A method of producing a poxvirus at an increased growth rate or titer, the method comprising:

contacting a host cell with a nucleocytoplasmic transport inhibitor at a concentration of at least 0.01 μM; wherein the host cell is a transformed cell;

contacting the host cell with a poxvirus, wherein the host cell is contacted with the poxvirus at a multiplicity of infection (MOI) less than one; and

culturing the host cell to produce progeny of the poxvirus at an increased growth rate compared to the growth rate in identical conditions lacking the nucleocytoplasmic transport inhibitor or an at least 3-fold increase in titer compared to that produced in identical conditions lacking the nucleocytoplasmic transport inhibitor,

wherein the nucleocytoplasmic transport inhibitor is one or more selected from the group consisting of: Leptomycin A, Leptomycin B, Ratjadone A, Ratjadone B, Ratjadone C, Ratjadone D, and Anguinomycin A; and

wherein the poxvirus is myxoma virus.

2. The method of claim 1 , further comprising harvesting the progeny of the poxvirus.

3. The method of claim 1 , wherein the host cell is contacted with the nucleocytoplasmic transport inhibitor prior to contacting the host cell with the poxvirus.

4. The method of claim 1 , wherein the host cell is contacted with the nucleocytoplasmic transport inhibitor after contacting the host cell with the poxvirus.

5. The method of claim 1 , wherein the host cell is contacted with the nucleocytoplasmic transport inhibitor and the poxvirus at the same time.

6. The method of claim 1 , wherein the poxvirus is genetically modified.

7. The method of claim 1 , wherein the culturing is conducted under good manufacturing practices (GMP) for manufacture of the poxvirus.

8. The method of claim 1 , wherein the host cell is contacted with the poxvirus at a multiplicity of infection (MOI) of between about 0.01 and about 0.1.

9. The method of claim 1 , wherein the increase in viral titer is evaluated 24 hours or 48 hours after contacting the host cell with the poxvirus.

10. A method for replicating a leporipoxvirus, the method comprising:

exposing a host cell to a nucleocytoplasmic transport inhibitor at a concentration of at least 0.01 μM; wherein the host cell is a transformed cell;

infecting the host cell with a leporipoxvirus at a multiplicity of infection (MOI) less than 1, thereby generating an infected host cell; and

incubating the infected host cell in a culture medium under conditions that allow for replication of the leporipoxvirus,

wherein the rate of replication is greater than the rate of replication in identical conditions lacking the nucleocytoplasmic transport inhibitor or an at least 3-fold increase in titer compared to that produced in identical conditions lacking the nucleocytoplasmic transport inhibitor

wherein the nucleocytoplasmic transport inhibitor is one or more selected from the group consisting of: Leptomycin A, Leptomycin B, Ratjadone A, Ratjadone B, Ratjadone C, Ratjadone D, and Anguinomycin A; and

wherein the leporipoxvirus is a myxoma virus.

11. The method of claim 1 , wherein the host cell is A549 cell.

12. The method of claim 10 , wherein the host cell is A549 cell.

Assignments (2)
CONFIRMATORY LICENSE Recorded Oct 12, 2022
From: ARIZONA STATE UNIVERSITY, TEMPE CAMPUS
To: NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF HEALTH AND HUMAN SERVICES (DHHS), U.S. GOVERNMENT
Reel/Frame 061661/0339 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Sep 22, 2021
From: MCFADDEN, DOUGLAS GRANT; RAHMAN, MOHAMMED MASMUDUR
To: ARIZONA BOARD OF REGENTS ON BEHALF OF ARIZONA STATE UNIVERSITY
Reel/Frame 057565/0455 →
Continuity (2)
Provisional Application 62716224 · Aug 8, 2018
Related Publication 20210301263A1 · Sep 30, 2021
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