快速检测基因治疗候选者中腺相关病毒8型结合抗体：即时检测方法的开发

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

作者信息

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

机构信息

Chembio Diagnostics Inc., Research & Development, Medford, NY, USA.

Takeda Pharmaceutical Company Limited, Diagnostic Devices Center of Excellence, Cambridge, MA, USA.

出版信息

Gene Ther. 2025 Aug 21. doi: 10.1038/s41434-025-00559-0.

DOI:10.1038/s41434-025-00559-0

PMID:40841775

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.

摘要

预先存在的抗腺相关病毒（AAV）抗体对腺相关病毒（AAV）介导的基因治疗的成功构成了重大挑战，因为它们会降低治疗效果，限制患者的治疗资格，并在治疗过程中引发严重的健康问题。本研究引入了首个即时检测（POC）测试，用于快速、定量检测患者血浆、血清和血液中的AAV8结合抗体，该测试利用了Chembio的双路径平台（DPP）技术。DPP AAV8总抗体（TAb）检测在加入样品后20分钟内即可得出结果，在用与AAV8结合的纯化人多克隆抗体进行评估时，动态范围为0-32μg/ml，相对于Chembio的AAV8 TAb ELISA具有合理的特异性和灵敏度（R² = 0.90）。此外，该检测与Chembio的AAV8中和抗体（NAb）ELISA和基于细胞的NAb检测显示出很强的相关性（血浆中R² = 0.97）（基于细胞的检测改编自BioAgilytix欧盟协议）。这种快速可靠的检测可以促进对潜在基因治疗患者进行筛查，以检测其是否预先存在与AAV8结合的抗体，并评估他们是否适合接受AAV8介导的基因治疗。