TY - JOUR
T1 - AAV-ID
T2 - A Rapid and Robust Assay for Batch-to-Batch Consistency Evaluation of AAV Preparations
AU - Pacouret, Simon
AU - Bouzelha, Mohammed
AU - Shelke, Rajani
AU - Andres-Mateos, Eva
AU - Xiao, Ru
AU - Maurer, Anna
AU - Mevel, Mathieu
AU - Turunen, Heikki
AU - Barungi, Trisha
AU - Penaud-Budloo, Magalie
AU - Broucque, Frédéric
AU - Blouin, Véronique
AU - Moullier, Philippe
AU - Ayuso, Eduard
AU - Vandenberghe, Luk H.
N1 - Publisher Copyright:
© 2017 The American Society of Gene and Cell Therapy
PY - 2017/6/7
Y1 - 2017/6/7
N2 - 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.
AB - 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.
KW - AAV vectors
KW - CMC
KW - adeno-associated virus
KW - capsid thermostability
KW - gene therapy
KW - homogeneity
KW - identity
KW - identity assay
KW - manufacturing
KW - quality control
UR - https://www.scopus.com/pages/publications/85017434235
U2 - 10.1016/j.ymthe.2017.04.001
DO - 10.1016/j.ymthe.2017.04.001
M3 - Article
C2 - 28427840
AN - SCOPUS:85017434235
SN - 1525-0016
VL - 25
SP - 1375
EP - 1386
JO - Molecular Therapy
JF - Molecular Therapy
IS - 6
ER -
