To ensure the safety and efficacy of viral-vector-based gene therapy products, robust and reliable characterization tools are essential across development, production and quality control.
Light-scattering-based analytical techniques for AAV analysis quantify key attributes including size, aggregation, physical titer, empty:full ratio and stability.
Learn how Wyatt’s solutions for characterizing critical quality attributes of AAVs and other DNA/RNA delivery modalities meet the needs of industry and regulators. They are simple to operate and provide high productivity, yet directly access the core biophysical properties that define these CQAs.
SEC-MALS combines separation by size-exclusion chromatography with analysis by multi-angle light scattering, refractive index and UV detectors. Quantifying multiple AAV CQAs by SEC-MALS is fast, accurate, automated and highly repeatable, requiring just a single HPLC run.
The method uses a DAWN® MALS instrument, an Optilab® refractive index detector and a HPLC UV module to measure size, titer (capsids/mL), aggregated fraction, and relative capsid content (empty:full ratio) of the vectors. Results compare favorably with expected values and those obtained by traditional, more laborious methods. Details are available in application note AN1617: Quantifying quality attributes of AAV gene therapy vectors by SEC-UV-MALS-dRI.
Dynamic light scattering performs rapid, low-volume measurements in quartz cuvettes or microwell plates. DLS is optimal for determining multiple AAV properties over a series of formulation or process conditions, and for assessing stability over temperature or time courses.
The method uses a DynaPro® NanoStar® cuvette-based DLS instrument for testing only few samples, or a DynaPro® Plate Reader when many samples, conditions and replicates are required. The results include size, size distributions and titer (capsids/mL).
Results compare favorably with those obtained by SEC-MALS (see above). Changes in size in the course of a temperature ramp indicate stability of the capsid and constituent proteins. Details are available in application note AN5007: Characterization of AAV-based viral vectors by DynaPro DLS/SLS instruments.