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Author Notes:

Matthew R. Gardner, Division of Infectious Diseases, Department of Medicine, Emory University, Atlanta, GA 30329, USA. Email: matthew.r.gardner@emory.edu

M.R.G. designed the study and performed experiments. D.E.M. and C.P.M. performed experiments. J.M.M.-N. provided critical samples for analysis. S.P.F. performed data analysis. G.G. provided critical reagents. R.C.D. designed and supervised the study. M.R.G. and R.C.D. wrote the manuscript with input and approval from all the coauthors.

We would like to the thank William Lauer for his technical expertise and coordinating of sample distribution; Eva Rakasz for rhesus macaque sample collection and distribution; and Meredith E. Davis-Gardner for her insights and comments on the manuscript. The graphical abstract was created using BioRender.com.

M.R.G. is a co-founder and consultant for Emmune, Inc. M.R.G. is an inventor on patents with potential royalties licensed to Emmune, Inc. G.G. is a co-founder of Voyager Therapeutics and Aspa Therapeutics and holds equity in both companies. G.G. is an inventor on patents with potential royalties licensed to Voyager Therapeutics, Aspa Therapeutics, and other biopharmaceutical companies. The remaining authors declare no competing interests.

Subjects:

Research Funding:

This work was supported by the Emory University Molecules and Pathogens to Populations and Pandemics (MP3) Initiative and by NIH grants R00AI138860 (M.R.G.), R01AI098446 (R.C.D.), and U19AI149646 (R.C.D.).

Keywords:

  • Science & Technology
  • Life Sciences & Biomedicine
  • Medicine, Research & Experimental
  • Research & Experimental Medicine
  • ADENOASSOCIATED VIRUS
  • IMMUNE-RESPONSES
  • PREVALENCE
  • CHILDREN
  • DELIVERY

High concordance of ELISA and neutralization assays allows for the detection of antibodies to individual AAV serotypes

Tools:

Journal Title:

MOLECULAR THERAPY-METHODS & CLINICAL DEVELOPMENT

Volume:

Volume 24

Publisher:

, Pages 199-206

Type of Work:

Article | Final Publisher PDF

Abstract:

Prescreening of participants in clinical trials that use adeno-associated virus (AAV) vectors is required to identify naive participants, as preexisting neutralizing antibodies can limit the efficacy of AAV gene therapies. The presence of antibodies to individual AAV serotypes is typically detected by neutralization assay. To streamline the screening process, we compared an ELISA-based screening method with a neutralization assay for the detection of antibodies against AAV1, AAV8, and AAV9 in a collection of 50 rhesus macaque sera and 20 human sera. We observed a high level of concordance between the two assays (Pearson r > 0.8) for all three serotypes in both sample sets. We thus investigated pre- vs post-vector inoculation sera samples from rhesus macaques that received AAV1 or AAV8 vector inoculations for cross-reactive anti-AAV antibodies. All 12 macaques seroconverted to the vector they received, but many also reacted to the other serotypes. Our results validate an easy-to-use ELISA for reliable detection of antibodies to individual serotypes of AAV. Our results also demonstrate that an antibody response post-AAV inoculation may partially cross-react with other AAV serotypes. Overall, these results suggest that either assay can be used by academic labs for prescreening samples for preexisting anti-AAV antibodies.

Copyright information:

© 2022 The Authors

This is an Open Access work distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (https://creativecommons.org/licenses/by-nc-nd/4.0/rdf).
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