Stunner AAV / AdV 病毒載體快速定量分析

Stunner from Unchained Labs is the only plate-based platform that combines UV/Vis spectroscopy, dynamic light scattering (DLS), and static light scattering (SLS) to deliver comprehensive AAV and AdV characterization from just 2 µL of sample. This innovative system eliminates the need for traditional qPCR and ELISA workflows by providing simultaneous measurement of capsid titer, empty/full ratio, and aggregation detection in a single, streamlined assay.

For AAV (adeno-associated virus) analysis, Stunner utilizes the AAV Quant application that integrates UV/Vis absorbance data with DLS and SLS measurements. The system accurately quantifies total capsid titer (cp/mL), full capsid titer (vg/mL), free and aggregated protein, and free and aggregated ssDNA. The Unmix algorithm automatically deconvolutes the absorbance spectrum to separate protein and nucleic acid contributions, eliminating the need for manual extinction coefficient calculations or concerns about overlapping spectra.

Stunner supports multiple AAV serotypes including AAV2, AAV5, AAV8, and AAV9, with preloaded parameters for each serotype. Users can also input custom viral genome sequences and extinction coefficients for novel AAV variants. The system delivers empty/full ratio measurements with impressive precision, typically achieving standard deviations of less than 5% for known mixtures of empty and full capsids.

For larger viral vectors such as adenovirus (AdV), Stunner employs rotating angle dynamic light scattering (RADLS) and multi-angle light scattering (MALS) technology. The Adeno Quant application provides accurate particle counting for larger vectors, with a detection range extending down to 10^9 cp/mL. RADLS enables precise quantification of AdV total capsid titer, full capsid titer, and empty/full ratio without interference from aggregates or residual DNA contaminants.

The platform features a unique microfluidic plate design with 96 sample positions in standard SBS format. Each microfluidic circuit incorporates two fixed pathlengths (0.1 mm and 0.7 mm) to cover a wide dynamic range of 0.03-275 OD. A full 96-well plate can be measured in approximately 1 hour for AAV Quant or 2 hours 15 minutes for Adeno Quant applications. The system is automation-friendly and can be integrated with robotic liquid handlers for high-throughput workflows.

DLS analysis on Stunner enables rapid detection of aggregates that can compromise downstream assays and therapeutic efficacy. The intensity distribution clearly displays the relative scattering from intact capsids versus aggregated material, allowing researchers to quickly identify problematic samples before investing time in cell-based assays. This capability is particularly valuable for formulation screening and stability studies, where buffer conditions and storage parameters significantly impact AAV aggregation propensity.

Stunner's dye-free, label-free, and standard-free workflow significantly reduces assay development time compared to traditional methods. The system requires no sample preparation or dilution, enabling direct measurement of samples across a broad concentration range. This streamlined approach makes it possible to check capsid titer and empty/full ratio more frequently throughout process development and manufacturing.

For regulatory environments, Stunner offers an optional 21 CFR Part 11 compliant software package and supports USP and Ph. Eur. performance verification standards. The instrument specifications include CE, FCC, and CSA approvals, making it suitable for both research and GMP-adjacent applications.

Key performance specifications include wavelength range of 230-750 nm for UV/Vis measurements, size accuracy of ±2% for DLS, and hydrodynamic diameter range of 0.3-1000 nm. The system achieves absorbance precision of ±0.01 OD (standard deviation) for samples below 1 OD and ±1% CV for samples in the 1-200 OD range.

Stunner represents a significant advancement in viral vector analytics, providing gene therapy researchers and manufacturers with rapid, accurate, and comprehensive characterization data from minimal sample volumes. By combining multiple analytical techniques in a single platform, Stunner accelerates AAV and AdV development workflows while ensuring consistent quality throughout the production process.