Development of a Robust Downstream Strategy for High Quality AAV Using an All-column Purification Method
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Abstract
Andelyn Biosciences has recently developed a robust scalable suspension platform for production of high yields of AAV for preclinical and all phases of clinical production. This platform is based on a quality by design approach that ensures scalability and quality at different stages of development. The standard purification strategy involves use of ultracentrifugation for enrichment of full capsids. An alternative all-column chromatography technique will be presented which replaces ultracentrifugation for downstream purification of AAV. The process flow includes concentration of the clarified harvest followed by two IEX chromatography steps – one for capture and another for enrichment of full capsids. Data will be presented showing reproducibility and robustness of the strategy, with consistency in recovery and purity at each stage. Compared to the standard ultracentrifugation approach for enriching full capsids, the all-column method results in 75% reduction in processing time. The IEX procedure results in 50% recovery which consists of 88-98% full capsids with final purity of 99-100% as evaluated using HPLC, AUC and capillary electrophoresis.
Download our ESGCT 2022 Poster
Get exclusive access to our ESGCT 2022 poster! Enter your email to download the poster and stay informed on how we’re pushing the boundaries of cell and gene therapy for a better future.
Abstract
Andelyn Biosciences has recently developed a robust scalable suspension platform for production of high yields of AAV for preclinical and all phases of clinical production. This platform is based on a quality by design approach that ensures scalability and quality at different stages of development. The standard purification strategy involves use of ultracentrifugation for enrichment of full capsids. An alternative all-column chromatography technique will be presented which replaces ultracentrifugation for downstream purification of AAV. The process flow includes concentration of the clarified harvest followed by two IEX chromatography steps – one for capture and another for enrichment of full capsids. Data will be presented showing reproducibility and robustness of the strategy, with consistency in recovery and purity at each stage. Compared to the standard ultracentrifugation approach for enriching full capsids, the all-column method results in 75% reduction in processing time. The IEX procedure results in 50% recovery which consists of 88-98% full capsids with final purity of 99-100% as evaluated using HPLC, AUC and capillary electrophoresis.
Download our ESGCT 2022 Poster
Get exclusive access to our ESGCT 2022 poster! Enter your email to download the poster and stay informed on how we’re pushing the boundaries of cell and gene therapy for a better future.
Abstract
Andelyn Biosciences has recently developed a robust scalable suspension platform for production of high yields of AAV for preclinical and all phases of clinical production. This platform is based on a quality by design approach that ensures scalability and quality at different stages of development. The standard purification strategy involves use of ultracentrifugation for enrichment of full capsids. An alternative all-column chromatography technique will be presented which replaces ultracentrifugation for downstream purification of AAV. The process flow includes concentration of the clarified harvest followed by two IEX chromatography steps – one for capture and another for enrichment of full capsids. Data will be presented showing reproducibility and robustness of the strategy, with consistency in recovery and purity at each stage. Compared to the standard ultracentrifugation approach for enriching full capsids, the all-column method results in 75% reduction in processing time. The IEX procedure results in 50% recovery which consists of 88-98% full capsids with final purity of 99-100% as evaluated using HPLC, AUC and capillary electrophoresis.