Kovács Beatrix, Somogyi Fanni, Szabó Viktória, Nagy Zoltán Zsolt, Hernádi István, Mátyás Ferenc, Vanduffel Wim, Szemlaky Zsuzsanna, Rózsa Balázs, Ulbert István, Hillier Dániel
Institute of Cognitive Neuroscience and Psychology, HUN-REN Research Centre for Natural Sciences, Budapest, Hungary.
János Szentágothai Neurosciences Division, Doctoral School, Semmelweis University, Budapest, Hungary.
Front Immunol. 2025 Aug 20;16:1623848. doi: 10.3389/fimmu.2025.1623848. eCollection 2025.
Adeno-associated virus (AAV) gene therapy is often limited by pre-existing neutralizing antibodies (NAbs), yet current assays for NAb detection lack standardization and rarely quantify uncertainty, complicating cross-study comparisons. We present coreTIA (core Transduction Inhibition Assay), a comprehensive framework providing a modular experimental protocol and a statistically robust analysis pipeline. This integrated method delivers precise, reproducible NAb titers with quantified uncertainty for every result. coreTIA's statistical framework enables robust estimation of neutralization even when dilution series are incomplete, helping to reduce repeat testing and minimizing sample volume requirements. Evaluation and refinement of key assay parameters support consistent performance across AAV serotypes. By providing a protocol and analysis suite as an open resource, coreTIA facilitates more consistent and transparent NAb measurement, potentially aiding assay harmonization and regulatory assessment, addressing a key barrier to progress in gene therapy research and development.
腺相关病毒（AAV）基因疗法常常受到预先存在的中和抗体（NAb）的限制，然而目前用于检测NAb的检测方法缺乏标准化，而且很少对不确定性进行量化，这使得跨研究比较变得复杂。我们提出了coreTIA（核心转导抑制试验），这是一个全面的框架，提供了模块化的实验方案和具有统计学稳健性的分析流程。这种综合方法为每个结果提供了精确、可重复的NAb滴度，并对不确定性进行了量化。coreTIA的统计框架即使在稀释系列不完整时也能实现对中和作用的稳健估计，有助于减少重复检测并将样本量要求降至最低。对关键检测参数的评估和优化支持了AAV各血清型之间的一致性能。通过将方案和分析套件作为开放资源提供，coreTIA促进了更一致、更透明的NAb测量，有可能有助于检测方法的协调和监管评估，解决了基因治疗研发进展的一个关键障碍。
