PCA Raman Analysis of AAV Serotypes and Full vs Empty Particles

The patent replaces the mechanical and biochemical processes of droplet digital PCR with a non-invasive optical detection method using Raman spectroscopy. The system uses laser excitation to generate Raman signals from the viral particles, which are then detected and analyzed to determine genome titration, eliminating the need for complex PCR machinery and reagents

The Raman spectroscopy system performs self-service by directly detecting the intrinsic vibrational modes of molecular bonds in the viral particles without requiring external labels, tags, or complex sample preparation. The method uses the natural Raman scattering properties of the viral components to obtain analytical information

The patent transitions from one-dimensional intensity-based Raman analysis to multi-dimensional spectral analysis by examining multiple wavenumber ranges simultaneously. The system analyzes specific spectral regions (e.g., 489-728 cm⁻¹ for nucleic acids, 1645-1680 cm⁻¹ for proteins) to extract multiple attributes from a single Raman spectrum, enabling serotype differentiation and loading status determination without ligand binding

The patent changes the analytical parameters by using chemometric and statistical data analysis approaches to process the Raman spectral data. Methods such as principal component analysis (PCA), partial least squares (PLS), and other multivariate analysis techniques transform the raw spectral data into meaningful parameters that differentiate between serotypes and loading statuses