Size-exclusion chromatography as a multi-attribute method for process and product characterization of adeno-associated virus
Adeno-associated viruses (AAVs) have recently emerged as a leading platform for gene therapy. Due to the complex manufacturing process and structural features of AAVs, extensive process and product characterization studies are required to better understand product quality and batch-to-batch variabil...
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| Format: | Article |
| Language: | English |
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Elsevier
2024-12-01
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| Series: | Molecular Therapy: Methods & Clinical Development |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2329050124001980 |
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| author | Sri Hari Raju Mulagapati Arun Parupudi Tomasz Witkos Nick Bond Xiaoyu Chen Thomas Linke Guoling Xi Albert Ethan Schmelzer Wei Xu |
| author_facet | Sri Hari Raju Mulagapati Arun Parupudi Tomasz Witkos Nick Bond Xiaoyu Chen Thomas Linke Guoling Xi Albert Ethan Schmelzer Wei Xu |
| author_sort | Sri Hari Raju Mulagapati |
| collection | DOAJ |
| description | Adeno-associated viruses (AAVs) have recently emerged as a leading platform for gene therapy. Due to the complex manufacturing process and structural features of AAVs, extensive process and product characterization studies are required to better understand product quality and batch-to-batch variability. It is, therefore, critical to develop a fast and reliable analytical method to monitor different product quality attributes (PQAs) of AAVs. In this study, we developed a multiple-attribute monitoring (MAM) method for the characterization of AAV PQAs. The MAM method was developed using the separation capability of size-exclusion chromatography (SEC) in connection with multiple in-line detectors: ultraviolet (UV), fluorescence (FLD), multi-angle light scattering (MALS), and refractive index (RI). We demonstrate that our SEC-based MAM method can be used to measure different PQAs, including genome and capsid titer, purity, aggregation, and full/empty capsid ratios in a single assay. Our SEC-based MAM method achieves similar results when compared side by side with orthogonal, individual assays such as quantitative polymerase chain reaction (qPCR), enzyme-linked immunosorbent assay (ELISA), and anion-exchange chromatography (AEX). Moreover, here we demonstrate that a simple, label-free, cost-effective, minimum sample requirement, and a high-throughput method can be applied to support process development, product characterization, release, and stability testing. |
| format | Article |
| id | doaj-art-19f3cc3d9150449a9cb45f7162a32244 |
| institution | Kabale University |
| issn | 2329-0501 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Methods & Clinical Development |
| spelling | doaj-art-19f3cc3d9150449a9cb45f7162a322442024-12-03T04:28:52ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012024-12-01324101382Size-exclusion chromatography as a multi-attribute method for process and product characterization of adeno-associated virusSri Hari Raju Mulagapati0Arun Parupudi1Tomasz Witkos2Nick Bond3Xiaoyu Chen4Thomas Linke5Guoling Xi6Albert Ethan Schmelzer7Wei Xu8Process and Analytical Sciences, BioPharmaceuticals Development (BPD), R&D, AstraZeneca, Gaithersburg, MD, USA; Corresponding author: Sri Hari Raju Mulagapati, Process and Analytical Sciences, BioPharmaceuticals Development (BPD), R&D, AstraZeneca, Gaithersburg, MD, USA.Drug Product and Formulation Sciences, GSK Vaccines, Rockville, MD 20850, USAProcess and Analytical Sciences, BioPharmaceuticals Development (BPD), R&D, AstraZeneca, Gaithersburg, MD, USAProcess and Analytical Sciences, BioPharmaceuticals Development (BPD), R&D, AstraZeneca, Gaithersburg, MD, USAProcess and Analytical Sciences, BioPharmaceuticals Development (BPD), R&D, AstraZeneca, Gaithersburg, MD, USAProcess and Analytical Sciences, BioPharmaceuticals Development (BPD), R&D, AstraZeneca, Gaithersburg, MD, USAProcess and Analytical Sciences, BioPharmaceuticals Development (BPD), R&D, AstraZeneca, Gaithersburg, MD, USAViral Vector Product Development, Process Development and Clinical Supply, Alexion, Boston, MA, USAProcess and Analytical Sciences, BioPharmaceuticals Development (BPD), R&D, AstraZeneca, Gaithersburg, MD, USAAdeno-associated viruses (AAVs) have recently emerged as a leading platform for gene therapy. Due to the complex manufacturing process and structural features of AAVs, extensive process and product characterization studies are required to better understand product quality and batch-to-batch variability. It is, therefore, critical to develop a fast and reliable analytical method to monitor different product quality attributes (PQAs) of AAVs. In this study, we developed a multiple-attribute monitoring (MAM) method for the characterization of AAV PQAs. The MAM method was developed using the separation capability of size-exclusion chromatography (SEC) in connection with multiple in-line detectors: ultraviolet (UV), fluorescence (FLD), multi-angle light scattering (MALS), and refractive index (RI). We demonstrate that our SEC-based MAM method can be used to measure different PQAs, including genome and capsid titer, purity, aggregation, and full/empty capsid ratios in a single assay. Our SEC-based MAM method achieves similar results when compared side by side with orthogonal, individual assays such as quantitative polymerase chain reaction (qPCR), enzyme-linked immunosorbent assay (ELISA), and anion-exchange chromatography (AEX). Moreover, here we demonstrate that a simple, label-free, cost-effective, minimum sample requirement, and a high-throughput method can be applied to support process development, product characterization, release, and stability testing.http://www.sciencedirect.com/science/article/pii/S2329050124001980size-exclusion chromatographymulti-attribute monitoringMAMadeno-associated virusesdifferent in-line detectorsproduct quality attributes |
| spellingShingle | Sri Hari Raju Mulagapati Arun Parupudi Tomasz Witkos Nick Bond Xiaoyu Chen Thomas Linke Guoling Xi Albert Ethan Schmelzer Wei Xu Size-exclusion chromatography as a multi-attribute method for process and product characterization of adeno-associated virus Molecular Therapy: Methods & Clinical Development size-exclusion chromatography multi-attribute monitoring MAM adeno-associated viruses different in-line detectors product quality attributes |
| title | Size-exclusion chromatography as a multi-attribute method for process and product characterization of adeno-associated virus |
| title_full | Size-exclusion chromatography as a multi-attribute method for process and product characterization of adeno-associated virus |
| title_fullStr | Size-exclusion chromatography as a multi-attribute method for process and product characterization of adeno-associated virus |
| title_full_unstemmed | Size-exclusion chromatography as a multi-attribute method for process and product characterization of adeno-associated virus |
| title_short | Size-exclusion chromatography as a multi-attribute method for process and product characterization of adeno-associated virus |
| title_sort | size exclusion chromatography as a multi attribute method for process and product characterization of adeno associated virus |
| topic | size-exclusion chromatography multi-attribute monitoring MAM adeno-associated viruses different in-line detectors product quality attributes |
| url | http://www.sciencedirect.com/science/article/pii/S2329050124001980 |
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