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. 2025 Aug 21.
doi: 10.1038/s41434-025-00559-0. Online ahead of print.

Rapid detection of AAV8 binding antibodies in gene therapy candidates: development of a point-of-care approach

Alexander Kozikowski #  1 , Qing Wang #  1 , Cheng Yang  2 , Neil Gordon  1 , Kristina M Ciociola  2 , Asanka Yapa  1 , Claudia Villa  1 , Paul Lambotte  1 , Thomas Pisani  2 , Javan Esfandiari  1 , Angelo H Gunasekera  3
Affiliations

Rapid detection of AAV8 binding antibodies in gene therapy candidates: development of a point-of-care approach

Alexander Kozikowski et al. Gene Ther. .

Abstract

Preexisting anti-AAV antibodies pose a significant challenge to the success of Adeno-associated Virus (AAV) mediated gene therapies, as they can diminish therapeutic effectiveness, restrict patient eligibility for treatment, and cause serious health issues during treatment. This study introduces the first point-of-care (POC) test for the rapid, quantitative detection of AAV8 binding antibodies in patients' plasma, serum, and blood, leveraging Chembio's Dual Path Platform (DPP) technology. The DPP AAV8 Total Antibody (TAb) assay delivers results within 20 min from sample addition, with a dynamic range of 0-32 µg/ml when evaluated with purified human polyclonal antibodies that bind to AAV8, with reasonable specificity and sensitivity relative to Chembio's AAV8 TAb ELISA (R² = 0.90). Moreover, the assay demonstrated strong correlations with Chembio's AAV8 neutralizing antibody (NAb) ELISA and cell-based NAb assays (R² = 0.97 in plasma) (Cell-based assay adapted from BioAgilytix EU protocol). This rapid and reliable test can facilitate the screening of potential gene therapy patients for pre-existing antibodies that bind to AAV8 and assess their suitability for AAV8-mediated gene therapy.

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Conflict of interest statement

Competing interests: AK, QW, NG, AY, CV, PL, JE, and AG are current or past employees of Chembio Diagnostics Inc. and have no financial interests or personal relationships that could have influenced the work reported in this paper. CY, KC, and TP are current or past employees of Takeda. Takeda provided financial support. The Takeda authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed. No writing assistance was utilized in the production of this manuscript. Ethics approval and consent to participate statement: All methods were carried out following relevant guidelines and regulations. The study did not involve any live vertebrate animals; therefore, no ethical approval or reference/registration number was required. All biological samples were obtained from commercial vendors and were fully de-identified, with no identifiable information linked to individuals. As no human research participants were involved in the study, informed consent was not required.

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