Human Gene Therapy. 2021 Apr 23
Hubert D.-J. Daniel, Sanjay Kumar, Rajesh Kannangai, Kavitha M. Lakshmi, Mavis Agbandje-Mckenna, Kirsten Coleman, Arun Srivastava, Alok Srivastava, and Asha Mary Abraham
Products used in the paper Details Operation
AAV vector packaging AAV3 vectors used for this study carried the gene coding for mCherry driven by the CMV promoter. The AAV3 was produced by using the triple- plasmidstransfection method and purified by using Iodixanol gradient ultra-centrifugation. The vector was procured from PackGene Biotech, LLC (Worcester, MA). Request Quote

Research Field: Hemophilia B

Keywords: AAV3, total capsid-binding antibody, neutralizing antibody, hemophilia B

AAV Serotype: AAV3

Dose: 10^4 VGs

Animal or cell line strain: COS-7 cells


Adeno-associated virus (AAV) vector-based gene therapy offers a new treatment option for individuals with hemophilia. Pre-existing anti-AAV antibodies significantly impact the use of AAV vectors. Even relatively low titers of AAV neutralizing antibodies (NAb) from natural AAV infections against the capsid have been shown to inhibit the transduction of intravenously administered AAV in animal models and were associated with limited efficacy in human trials. This is important for determining the primary eligibility of patients for AAV vector-based gene therapy clinical trials. Current techniques to screen AAV antibodies include AAV capsid enzyme-linked immunosorbent assay (ELISA) for total antibodies and a transduction inhibition assay (TIA) for NAb. This study developed and screened total capsid binding anti-AAV3 antibodies by using ELISA and determined NAb levels by TIA using mCherry flow cytometry in healthy individuals with hemophilia B in India. One hundred and forty-three apparently healthy controls and 92 individuals with hemophilia B were screened. The prevalence of total and NAb in healthy controls was 79.7% and 65%, respectively; the prevalence of total and NAb in patients with hemophilia B for AAV3 was 92.4% and 91.3%, respectively.

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