JPWO2019241535A5 - - Google Patents
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- JPWO2019241535A5 JPWO2019241535A5 JP2020568998A JP2020568998A JPWO2019241535A5 JP WO2019241535 A5 JPWO2019241535 A5 JP WO2019241535A5 JP 2020568998 A JP2020568998 A JP 2020568998A JP 2020568998 A JP2020568998 A JP 2020568998A JP WO2019241535 A5 JPWO2019241535 A5 JP WO2019241535A5
- Authority
- JP
- Japan
- Prior art keywords
- aav
- raav particles
- feed composition
- anion exchange
- exchange chromatography
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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- 241000702421 Dependoparvovirus Species 0.000 claims 32
- 239000002245 particle Substances 0.000 claims 26
- 238000000034 method Methods 0.000 claims 23
- 239000000203 mixture Substances 0.000 claims 18
- 239000012501 chromatography medium Substances 0.000 claims 17
- 238000005571 anion exchange chromatography Methods 0.000 claims 10
- 238000010828 elution Methods 0.000 claims 7
- 241000700605 Viruses Species 0.000 claims 6
- 239000011534 wash buffer Substances 0.000 claims 5
- 150000003839 salts Chemical class 0.000 claims 4
- 238000005406 washing Methods 0.000 claims 4
- 210000000234 capsid Anatomy 0.000 claims 3
- 108090000565 Capsid Proteins Proteins 0.000 claims 2
- 102100023321 Ceruloplasmin Human genes 0.000 claims 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims 2
- 239000012535 impurity Substances 0.000 claims 2
- 238000000926 separation method Methods 0.000 claims 2
- 241001655883 Adeno-associated virus - 1 Species 0.000 claims 1
- 241000702423 Adeno-associated virus - 2 Species 0.000 claims 1
- 241000202702 Adeno-associated virus - 3 Species 0.000 claims 1
- 241000580270 Adeno-associated virus - 4 Species 0.000 claims 1
- 241001634120 Adeno-associated virus - 5 Species 0.000 claims 1
- 241000972680 Adeno-associated virus - 6 Species 0.000 claims 1
- 241001164823 Adeno-associated virus - 7 Species 0.000 claims 1
- 241001164825 Adeno-associated virus - 8 Species 0.000 claims 1
- 241000649045 Adeno-associated virus 10 Species 0.000 claims 1
- 241000958487 Adeno-associated virus 3B Species 0.000 claims 1
- 102100035426 DnaJ homolog subfamily B member 7 Human genes 0.000 claims 1
- 101100285903 Drosophila melanogaster Hsc70-2 gene Proteins 0.000 claims 1
- 101100178718 Drosophila melanogaster Hsc70-4 gene Proteins 0.000 claims 1
- 101100178723 Drosophila melanogaster Hsc70-5 gene Proteins 0.000 claims 1
- 101000804114 Homo sapiens DnaJ homolog subfamily B member 7 Proteins 0.000 claims 1
- 101150090950 Hsc70-1 gene Proteins 0.000 claims 1
- 101100150366 Schizosaccharomyces pombe (strain 972 / ATCC 24843) sks2 gene Proteins 0.000 claims 1
- 125000003277 amino group Chemical group 0.000 claims 1
- 239000000872 buffer Substances 0.000 claims 1
- 239000011780 sodium chloride Substances 0.000 claims 1
Claims (19)
（ａ）ｒＡＡＶ粒子及びウイルス凝集体を含むフィード組成物を、アニオン交換クロマトグラフィー媒体に、前記ｒＡＡＶ粒子が前記クロマトグラフィー媒体に結合するのを可能にする条件下で接触させること、
（ｂ）直線勾配を用いて前記クロマトグラフィー媒体から前記ｒＡＡＶ粒子を溶出すること、ならびに
（ｃ）溶出された前記ｒＡＡＶ粒子を含む溶出液を回収すること
を含み、
以下：
ａ．前記フィード組成物が、約０．１ｍＭから約２０ｍＭの間のＭｇ２＋濃度を有すること、
ｂ．前記フィード組成物が、約０．１ｍＭから約２０ｍＭの間のＫ＋濃度を有すること、
ｃ．前記フィード組成物が、約６．５から約１０．５の間のｐＨを有すること、
によって特徴づけられる、前記方法。 A method for improving the separation of recombinant adeno-associated virus (rAAV) particles and virus aggregates during anion exchange chromatography.
(A) Bringing the feed composition containing the rAAV particles and virus aggregates into contact with the anion exchange chromatography medium under conditions that allow the rAAV particles to bind to the chromatography medium.
(B) eluting the rAAV particles from the chromatographic medium using a linear gradient, and (c) recovering the eluate containing the eluted rAAV particles.
Less than:
a. The feed composition has a Mg 2+ concentration between about 0.1 mM and about 20 mM.
b. The feed composition has a K + concentration between about 0.1 mM and about 20 mM.
c. The feed composition has a pH between about 6.5 and about 10.5.
Said method characterized by .
ｄ．前記溶出することが、０．１ＣＶ／分から５ＣＶ／分の間の流量で行われること、及び
ｅ．前記直線塩勾配が、約５から約１００ＣＶの間の体積を含むこと
によって特徴づけられる、請求項１に記載の方法。 Less than:
d . The elution is carried out at a flow rate between 0.1 CV / min and 5 CV / min, and e. The linear salt gradient comprises a volume between about 5 and about 100 CV.
The method of claim 1 , characterized by .
ａ．前記フィード組成物が、約０．５ｍＭから約１０ｍＭの間のＭｇ２＋濃度を有すること、
ｂ．前記フィード組成物が、約０．５ｍＭから約１０ｍＭの間のＫ＋濃度を有すること、及び
ｃ．前記フィード組成物が、約６．５から約１０．５の間のｐＨを有すること
によって特徴づけられる、請求項１に記載の方法。 Less than:
a. The feed composition has a Mg 2+ concentration between about 0.5 mM and about 10 mM.
b. The feed composition has a K + concentration between about 0.5 mM and about 10 mM, and c. The method of claim 1 , wherein the feed composition is characterized by having a pH between about 6.5 and about 10.5.
（ａ）前記フィード組成物を、アニオン交換クロマトグラフィー媒体に、前記ｒＡＡＶ粒子が前記クロマトグラフィー媒体に結合するのを可能にする条件下で接触させること、
（ｂ）直線勾配を用いて前記クロマトグラフィー媒体から前記ｒＡＡＶ粒子を溶出すること、及び
（ｃ）溶出された前記ｒＡＡＶ粒子を含む溶出液を回収すること
を含み、
（ｉ）前記不純物が、空のウイルスカプシド、充填不完全なウイルスカプシド、及び／またはウイルス凝集体を含み、
（ｉｉ）前記方法が、以下：
ａ．前記フィード組成物が、約０．１ｍＭから約２０ｍＭの間のＭｇ２＋濃度を有すること、
ｂ．前記フィード組成物が、約０．１ｍＭから約２０ｍＭの間のＫ＋濃度を有すること、
ｃ．前記フィード組成物が、約６．５から約１０．５の間のｐＨを有すること、
ｄ．前記溶出することが、０．１ＣＶ／分から５ＣＶ／分の間の流量で行われること、及び
ｅ．前記直線塩勾配が、約５から約１００ＣＶの間の体積を含むこと
のうちの１つ以上によって特徴づけられる、前記方法。 A method for isolating recombinant adeno-associated virus (rAAV) particles from a feed composition comprising rAAV particles and impurities.
(A) Contacting the feed composition with an anion exchange chromatography medium under conditions that allow the rAAV particles to bind to the chromatography medium.
It comprises (b) eluting the rAAV particles from the chromatographic medium using a linear gradient and (c) recovering the eluate containing the eluted rAAV particles.
(I) The impurities include empty virus capsids, incompletely filled virus capsids, and / or virus aggregates.
(Ii) The above method is as follows:
a. The feed composition has a Mg 2+ concentration between about 0.1 mM and about 20 mM.
b. The feed composition has a K + concentration between about 0.1 mM and about 20 mM.
c. The feed composition has a pH between about 6.5 and about 10.5.
d. The elution is carried out at a flow rate between 0.1 CV / min and 5 CV / min, and e. The method, wherein the linear salt gradient is characterized by one or more of comprising a volume between about 5 and about 100 CV.
（ａ）空のｒＡＡＶ粒子、充填不完全なｒＡＡＶ粒子、及び完全ｒＡＡＶ粒子を含むフィード組成物を、アニオン交換クロマトグラフィー媒体に、前記ｒＡＡＶ粒子が前記クロマトグラフィー媒体に結合するのを可能にする条件下で接触させること、
（ｂ）直線勾配を用いて前記クロマトグラフィー媒体から前記ｒＡＡＶ粒子を溶出すること、ならびに
（ｃ）溶出された前記ｒＡＡＶ粒子を含む溶出液を回収すること
を含み、
以下：
ａ．前記フィード組成物が、約０．１ｍＭから約２０ｍＭの間のＭｇ２＋濃度を有すること、
ｂ．前記フィード組成物が、約０．１ｍＭから約２０ｍＭの間のＫ＋濃度を有すること、
ｃ．前記フィード組成物が、約６．５から約１０．５の間のｐＨを有すること、
ｄ．前記溶出することが、０．１ＣＶ／分から５ＣＶ／分の間の流量で行われること、及び
ｅ．前記直線塩勾配が、約５から約１００ＣＶの間の体積を含むこと
のうちの１つ以上によって特徴づけられる、前記方法。 A method for improving the separation of empty or incompletely filled recombinant adeno-associated virus (rAAV) particles from complete rAAV particles during anion exchange chromatography.
(A) Conditions that allow a feed composition comprising empty rAAV particles, incompletely filled rAAV particles, and complete rAAV particles to bind to an anion exchange chromatography medium and the rAAV particles to the chromatography medium. Contact underneath,
(B) eluting the rAAV particles from the chromatographic medium using a linear gradient, and (c) recovering the eluate containing the eluted rAAV particles.
Less than:
a. The feed composition has a Mg 2+ concentration between about 0.1 mM and about 20 mM.
b. The feed composition has a K + concentration between about 0.1 mM and about 20 mM.
c. The feed composition has a pH between about 6.5 and about 10.5.
d. The elution is carried out at a flow rate between 0.1 CV / min and 5 CV / min, and e. The method, wherein the linear salt gradient is characterized by one or more of comprising a volume between about 5 and about 100 CV.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US201862684835P | 2018-06-14 | 2018-06-14 | |
| US62/684,835 | 2018-06-14 | ||
| PCT/US2019/037013 WO2019241535A2 (en) | 2018-06-14 | 2019-06-13 | Anion exchange chromatography for recombinant aav production |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| JP2021526829A JP2021526829A (en) | 2021-10-11 |
| JPWO2019241535A5 true JPWO2019241535A5 (en) | 2022-06-20 |
| JP7385603B2 JP7385603B2 (en) | 2023-11-22 |
Family
ID=67513723
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2020568998A Active JP7385603B2 (en) | 2018-06-14 | 2019-06-13 | Anion exchange chromatography for recombinant AAV production |
Country Status (9)
| Country | Link |
|---|---|
| US (1) | US20210370199A1 (en) |
| EP (1) | EP3807405A2 (en) |
| JP (1) | JP7385603B2 (en) |
| KR (1) | KR20210020100A (en) |
| CN (1) | CN112469822A (en) |
| AU (1) | AU2019285186A1 (en) |
| CA (1) | CA3102817A1 (en) |
| IL (1) | IL279193A (en) |
| WO (1) | WO2019241535A2 (en) |
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| CA3201743A1 (en) | 2020-12-16 | 2022-06-23 | Robert STADELMAN | Method of producing a recombinant adeno-associated virus particle |
| CN116848255A (en) | 2021-01-21 | 2023-10-03 | 再生生物股份有限公司 | Improved production of recombinant polypeptides and viruses |
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-
2019
- 2019-06-13 CN CN201980048303.1A patent/CN112469822A/en active Pending
- 2019-06-13 EP EP19748612.9A patent/EP3807405A2/en active Pending
- 2019-06-13 CA CA3102817A patent/CA3102817A1/en active Pending
- 2019-06-13 AU AU2019285186A patent/AU2019285186A1/en active Pending
- 2019-06-13 JP JP2020568998A patent/JP7385603B2/en active Active
- 2019-06-13 US US17/251,869 patent/US20210370199A1/en active Pending
- 2019-06-13 KR KR1020217000909A patent/KR20210020100A/en active Search and Examination
- 2019-06-13 WO PCT/US2019/037013 patent/WO2019241535A2/en unknown
-
2020
- 2020-12-03 IL IL279193A patent/IL279193A/en unknown
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