Nano-pave: a nanopore-based method for AAV capsid quality assessment - adeno-associated viruses (AAVS) are the leading vectors for gene therapy, valued for their safety, low immunogenicity, and…

The National Center for Advancing Translational Sciences awarded Goeppert LLC, a Philadelphia-based small disadvantaged business, $350,000 on July 1, 2026, under CFDA 93.350 (National Center for Advancing Translational Sciences) to develop NANO-PAVE, a nanopore-based platform for assessing adeno-associated virus (AAV) capsid quality in gene therapy manufacturing.

NANO-PAVE addresses a critical bottleneck in AAV clinical translation: current quality-control assays (ELISA, ddPCR, analytical ultracentrifugation) are costly, labor-intensive, serotype-dependent, and produce inconsistent results across laboratories and production runs. The technology will use solid-state nanopores to analyze individual AAV particles without labels, dyes, or antibodies, leveraging voltage-driven translocation and electro-deformation to distinguish full from empty capsids and measure capsid load across serotypes in real time. Phase I will establish feasibility by demonstrating robust particle discrimination, validating nanopore-derived measurements against orthogonal benchmarks, exploring how genome structure affects capsid mechanics, and testing multiple AAV serotypes to ensure broad applicability. Phase I deliverables include a validated, serotype-independent NANO-PAVE technology with optimized nanopore design and recapture protocol.

The award period runs from July 1, 2026, through June 30, 2027. Performance takes place in Philadelphia, Pennsylvania. This is a Project Grant, the assistance type for NCATS Phase I research feasibility studies.

Nano-pave: a nanopore-based method for AAV capsid quality assessment - adeno-associated viruses (AAVS) are the leading vectors for gene therapy, valued for their safety, low immunogenicity, and broad tissue tropism. However, large-scale clinical translation remains hindered by challenges in manufacturing quality control. A critical barrier is the inability to rapidly and reliably measure capsid load and the ratio of full to empty capsids-key attributes that directly affect therapeutic potency and safety. Current assays such as elisa (capsid titer), ddpcr (genome titer), and AUC (full/empty ratio) are costly, labor-intensive, and serotype-dependent, leading to inconsistent results across laboratories and production runs. To overcome these limitations, we will develop nano-pave (nanopore-based analysis of proportion of full and empty adeno-associated virus capsids via nanopore evaluation), a solid-state nanopore platform that analyzes individual AAV particles without labels, dyes, or antibodies. By leveraging voltage-driven translocation and electro-deformation, nano-pave directly links mechanical signatures to genome packaging state, enabling real-time, quantitative discrimination between full and empty capsids. In addition, its single-particle resolution will allow accurate measurement of capsid load across serotypes, providing a powerful complement to existing bulk assays. This approach is cost-effective, scalable, and adaptable to all serotypes, making IT ideally suited for integration into manufacturing pipelines. Phase I will establish feasibility by demonstrating robust particle discrimination, validating nanopore- derived measurements against orthogonal benchmarks, exploring genome-structure effects on capsid mechanics, and testing multiple AAV serotypes to ensure broad applicability of nano-pave across vector types. Phase I deliverables will include a validated serotype-independent nano-pave technology-integrating optimized nanopore design, recapture protocol, and machine-learning analysis-benchmarked against established assays to provide a rigorous, quantitative QC approach. This work will lay the foundation for phase II development of a scalable, in-process analytical tool capable of distinguishing therapeutically relevant capsids from product-related impurities (empty, partially filled capsids, aggregates) and process-related impurities such as host cell proteins and media components, enabling comprehensive quality assessment. Ultimately, nano-pave has the potential to become a transformative QC platform for academic, clinical, and industrial gene therapy programs, improving safety, efficacy, and manufacturing efficiency.