JP-2022539148-A5 -

Dates

Publication Date

20230525

Application Date

20200626

Description

All references cited herein, including publications, patent applications, and patents, are incorporated herein by reference to the same extent as if they were included in this book, each reference being individually and specifically cited as a reference. Finally, preferred embodiments of the present invention are described in separate sections. [Embodiment 1] A method for purifying adeno-associated virus (AAV), (a) A step of loading an AAV-containing solution onto an affinity resin targeting AAV at room temperature under conditions that allow bonding between the AAV in the solution and the affinity resin; (b) a step of performing at least one washing step at room temperature; and (c) A step of eluting AAV from affinity resin at a temperature below 18°C; Methods that include... [Embodiment 2] The method according to Embodiment 1, wherein the temperature in step (c) is 1°C to 12°C. [Embodiment 3] The method according to Embodiment 1, wherein the temperature in step (c) is 2°C to 8°C. [Embodiment 4] The method according to any one of embodiments 1 to 3, further comprising contacting the AAV-containing solution with an anion exchanger and eluting the AAV-containing solution from the anion exchanger before loading the AAV-containing solution into the affinity resin. [Embodiment 5] The method according to any one of embodiments 1 to 4, wherein at least two washing steps are performed at room temperature. [Embodiment 6] The method according to any one of embodiments 1 to 5, wherein at least three washing steps are performed at room temperature. [Embodiment 7] The method according to any one of embodiments 1 to 6, wherein at least four washing steps are performed at room temperature. [Embodiment 8] The method according to embodiment 5, wherein two cleaning steps are performed. [Embodiment 9] The method according to embodiment 6, wherein three cleaning steps are performed. [Embodiment 10] The method according to embodiment 7, wherein four cleaning steps are performed. [Method 11] The method according to any one of embodiments 5 to 10, wherein the cleaning step is performed continuously. [Embodiment 12] The method according to any one of Embodiments 1 to 11, wherein at least one washing buffer comprises about 10 mM to about 200 mM TrisHCl and about 50 mM to about 500 mM of the salt. [Embodiment 13] The method according to Embodiment 12, wherein at least one washing buffer comprises about 25 mM to about 100 mM TrisHCl and about 75 mM to about 250 mM of the salt. [Embodiment 14] The method according to Embodiment 13, wherein at least one washing buffer comprises about 40 mM to about 60 mM TrisHCl and about 100 mM to about 150 mM of the salt. [Embodiment 15] The method according to any one of embodiments 12 to 14, wherein at least one washing buffer has a pH of about 7.5 to about 9.2, about 8.0 to about 9.0, or about 8.0 to about 8.8. [Embodiment 16] The method according to Embodiment 15, wherein at least one washing buffer comprises about 50 mM TrisHCl and about 125 mM of the salt and has a pH of about 8.5. [Embodiment 17] The method according to any one of Embodiments 1 to 16, wherein at least one washing buffer comprises about 10 mM to about 2000 mM sodium acetate and about 0.001% to about 1% (w/w) of polysorbate 80. [Embodiment 18] The method according to Embodiment 17, wherein at least one washing buffer comprises about 50 mM to about 200 mM sodium acetate and about 0.005% to about 0.3% (w/w) of polysorbate 80. [Embodiment 19] The method according to Embodiment 18, wherein at least one washing buffer comprises about 90 mM to about 110 mM sodium acetate and about 0.05% to about 0.2% (w/w) of polysorbate 80. [Embodiment 20] The method according to any one of embodiments 17 to 19, wherein at least one washing buffer has a pH of about 5.0 to about 7.4, about 5.5 to about 7.0, or about 5.5 to about 6.5. [Embodiment 21] The method according to Embodiment 20, wherein at least one washing buffer comprises about 100 mM sodium acetate and about 0.1% (w/w) polysorbate 80 and has a pH of about 6.0. [Embodiment 22] The method according to any one of Embodiments 1 to 21, wherein at least one washing buffer comprises about 10 mM to about 200 mM TrisHCl and about 10% to about 75% (w/w) ethylene glycol. [Embodiment 23] The method according to Embodiment 22, wherein at least one washing buffer comprises about 25 mM to about 100 mM TrisHCl and about 25% to about 70% (w/w) ethylene glycol. [Embodiment 24] The method according to Embodiment 23, wherein at least one washing buffer comprises about 40 mM to about 60 mM TrisHCl and about 40% to about 60% (w/w) ethylene glycol. [Embodiment 25] The method according to any one of embodiments 22 to 24, wherein at least one washing buffer has a pH of about 7.5 to about 9.2, about 8.0 to about 9.0, or about 8.0 to about 8.8. [Embodiment 26] The method according to Embodiment 25, wherein at least one washing buffer comprises about 50 mM TrisHCl and about 50% (w/w) ethylene glycol and has a pH of