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AAV-ID: A Rapid and Robust Assay for Batch-to-Batch Consistency Evaluation of AAV Preparations
- Source :
- Molecular Therapy, Molecular Therapy, Cell Press, 2017, 25 (6), pp.1375-1386. ⟨10.1016/j.ymthe.2017.04.001⟩
- Publication Year :
- 2017
- Publisher :
- American Society of Gene & Cell Therapy, 2017.
-
Abstract
- International audience; Adeno-associated virus (AAV) vectors are promising clinical candidates for therapeutic gene transfer, and a number of AAV-based drugs may emerge on the market over the coming years. To insure the consistency in efficacy and safety of any drug vial that reaches the patient, regulatory agencies require extensive characterization of the final product. Identity is a key characteristic of a therapeutic product, as it ensures its proper labeling and batch-to-batch consistency. Currently, there is no facile, fast, and robust characterization assay enabling to probe the identity of AAV products at the protein level. Here, we investigated whether the thermostability of AAV particles could inform us on the composition of vector preparations. AAV-ID, an assay based on differential scanning fluorimetry (DSF), was evaluated in two AAV research laboratories for specificity, sensitivity, and reproducibility, for six different serotypes (AAV1, 2, 5, 6.2, 8, and 9), using 67 randomly selected AAV preparations. In addition to enabling discrimination of AAV serotypes based on their melting temperatures, the obtained fluorescent fingerprints also provided information on sample homogeneity, particle concentration, and buffer composition. Our data support the use of AAV-ID as a reproducible, fast, and low-cost method to ensure batch-to-batch consistency in manufacturing facilities and academic laboratories.
- Subjects :
- 0301 basic medicine
[SDV]Life Sciences [q-bio]
viruses
Genetic Vectors
Gene transfer
Computational biology
adeno-associated virus
Biology
medicine.disease_cause
identity assay
AAV vectors
03 medical and health sciences
Consistency (database systems)
Structure-Activity Relationship
Capsid
CMC
Drug Discovery
Genetics
medicine
Humans
quality control
Molecular Biology
Adeno-associated virus
identity
Pharmacology
capsid thermostability
Protein Stability
Protein level
Reproducibility of Results
Dependovirus
Virology
gene therapy
[SDV] Life Sciences [q-bio]
manufacturing
030104 developmental biology
homogeneity
Spectrometry, Fluorescence
Mutation
Molecular Medicine
Thermodynamics
Original Article
Capsid Proteins
Subjects
Details
- Language :
- English
- ISSN :
- 15250016 and 15250024
- Database :
- OpenAIRE
- Journal :
- Molecular Therapy, Molecular Therapy, Cell Press, 2017, 25 (6), pp.1375-1386. ⟨10.1016/j.ymthe.2017.04.001⟩
- Accession number :
- edsair.doi.dedup.....44e2a3d128dbf47191393d947a8e96e9