Anion exchange chromatography for recombinant AAV production
Inventors
ZHANG, Claire G. • Levine, Jeffrey • ZAFAR, Sara • GERNER, Franz M.
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-12070702-B2
Publication Date
2024-08-27
Expiration Date
Abstract
Provided herein are methods for the purification of recombinant Adeno-Associated Virus (rAAV) particles using anion exchange chromatography.
Core Innovation
The invention relates to a method for isolating recombinant adeno-associated virus (rAAV) particles from a feed composition containing rAAV particles and an impurity comprising empty viral capsid, partially filled viral capsid, and/or a viral aggregate. The method includes contacting the feed composition with an anion exchange chromatography media under conditions that allow binding of the rAAV particles to the chromatography media.
The method further includes eluting the rAAV particles from the chromatography media using a linear gradient and recovering an eluate comprising the eluted rAAV particles. The process is characterized by the feed having a Mg2+ concentration between about 0.1 mM and about 20 mM, a K+ concentration between about 0.1 mM and about 20 mM, and a pH between about 6.5 and about 10.5.
In additional aspects, the invention improves separation during anion exchange chromatography by applying the same characterization of the feed composition. Specifically, it improves the separation between empty or partially filled rAAV particles and full rAAV particles, and it also improves separation between rAAV particles and viral aggregates, using contacting, linear gradient elution, and recovery of an eluate comprising the eluted rAAV particles.
Claims Coverage
The partial content provides three independent claims, each characterized by anion exchange chromatography with linear gradient elution and recovery of an eluate comprising eluted rAAV, and each further characterized by feed Mg2+, feed K+, and feed pH ranges. Across the independent claims, the inventive scope covers isolating rAAV from a feed containing defined impurities, improving separation between empty or partially filled rAAV and full rAAV, and improving separation between rAAV particles and viral aggregates.
Ion-controlled anion exchange isolation of rAAV from feeds containing empty/partially filled capsids and/or aggregates
A method for isolating recombinant adeno-associated virus (rAAV) particles from a feed composition containing rAAV particles and an impurity comprising empty viral capsid, partially filled viral capsid, and/or a viral aggregate by contacting the feed composition with an anion exchange chromatography media under conditions that allow binding of the rAAV particles, eluting the rAAV particles using a linear gradient, and recovering an eluate comprising the eluted rAAV particles; characterized by the feed composition having a Mg2+ concentration between about 0.1 mM and about 20 mM, a K+ concentration between about 0.1 mM and about 20 mM, and a pH between about 6.5 and about 10.5.
Ion-controlled anion exchange separation of empty/partially filled rAAV from full rAAV
A method for improving the separation between empty or partially filled recombinant adeno-associated virus (rAAV) particles and full rAAV particles during anion exchange chromatography by contacting a feed composition containing empty, partially filled and full rAAV particles with an anion exchange chromatography media under conditions that allow binding of the rAAV particles, eluting the rAAV particles using a linear gradient, and recovering an eluate comprising the eluted rAAV particles; characterized by the feed composition having a Mg2+ concentration between about 0.1 mM and about 20 mM, a K+ concentration between about 0.1 mM and about 20 mM, and a pH between about 6.5 and about 10.5.
Ion-controlled anion exchange separation of rAAV from viral aggregates
A method for improving the separation between recombinant adeno-associated virus (rAAV) particles and viral aggregates during anion exchange chromatography by contacting a feed composition containing rAAV particles and viral aggregates with an anion exchange chromatography media under conditions that allow binding of the rAAV particles, eluting the rAAV particles using a linear gradient, and recovering an eluate comprising the eluted rAAV particles; characterized by the feed composition having a Mg2+ concentration between about 0.1 mM and about 20 mM, a K+ concentration between about 0.1 mM and about 20 mM, and a pH between about 6.5 and about 10.5.
Across the independent claims, the claim coverage centers on anion exchange chromatography in which rAAV binds to anion exchange chromatography media, elution is performed with a linear gradient, and the eluate containing the eluted rAAV is recovered, with separation or isolation characterized by feed Mg2+, feed K+, and feed pH ranges.
Stated Advantages
Improved separation between empty or partially filled rAAV particles and full rAAV particles during anion exchange chromatography.
Improved separation between rAAV particles and viral aggregates during anion exchange chromatography.
Increased yield as reported in examples (GC step yield improvements up to ~15% and higher).
Documented Applications
GMP-compliant rAAV purification using anion exchange chromatography for isolating rAAV particles from feeds containing empty/partially filled capsids and/or viral aggregates, including improved separation of full versus empty/partially filled particles and improved separation of full rAAV versus aggregates.
Interested in licensing this patent?