biostudies-literatureUnknown31Meierrieks FAdeno-associated virus (AAV) vectors are among the most prominent viral vectors for <i>in vivo</i> gene therapy, and their investigation and development using high-throughput techniques have gained increasing interest. However, sample throughput remains a bottleneck in most analytical assays. In this study, we compared commonly used analytical methods for AAV genome titer, capsid titer, and transducing titer determination with advanced methods using AAV2, AAV5, and AAV8 as representative examples. For the determination of genomic titers, we evaluated the suitability of qPCR and four different digital PCR methods and assessed the respective advantages and limitations of each method. We found that both ELISA and bio-layer interferometry provide comparable capsid titers, with bio-layer interfMolecular therapy. Methods & clinical development101118https://www.ebi.ac.uk/biostudies/studies/S-EPMC10562196biostudies-literatureUnveiling the secrets of adeno-associated virus: novel high-throughput approaches for the quantification of multiple serotypes.PMC10562196Patz MWolff MWPickl AKour AMeierrieks FPflanz KfalseUnveiling the secrets of adeno-associated virus: novel high-throughput approaches for the quantification of multiple serotypes.Adeno-associated virus (AAV) vectors are among the most prominent viral vectors for <i>in vivo</i> gene therapy, and their investigation and development using high-throughput techniques have gained increasing interest. However, sample throughput remains a bottleneck in most analytical assays. In this study, we compared commonly used analytical methods for AAV genome titer, capsid titer, and transducing titer determination with advanced methods using AAV2, AAV5, and AAV8 as representative examples. For the determination of genomic titers, we evaluated the suitability of qPCR and four different digital PCR methods and assessed the respective advantages and limitations of each method. We found that both ELISA and bio-layer interferometry provide comparable capsid titers, with bio-layer interf2023-01-01T00:00:00Z2023 Dec2025-04-05T12:06:15.783Z2025-04-05T12:06:15.783ZS-EPMC105621963782271710.1016/j.omtm.2023.101118