| Anion exchange chromatography |
Surface charge density |
High accuracy, complex workflow, medium turnaround time (30
min/sample), difficult to establish optimal method applicable to all
rAAV serotypes |
| Optical Density |
UV absorbance (density analysis) |
Fast turnaround time
(15 min/sample), requires highly purified
AAV to minimize interference with UV absorbance, varying concentration
requirements (5 × 1011–1 × 1013 GC/mL) |
| Transmission electron microscopy |
Image
analysis |
Direct characterization method, statistically
small sample
image size, high coefficient of variation, time-consuming (6 h/sample) |
| Charge detection
mass spectrometry |
Mass to charge ratio |
High accuracy, requires extensive preparation, time-consuming
(2 h/sample), low throughput, not easily accessible |
| Size-exclusion chromatography, multiangle light scattering |
Size exclusion and static light scattering |
High
accuracy, medium turnaround time (20–30 min/sample) requires long column equilibration times, relatively
high volumes of sample (30–50 μL per run), high concentration requirements (1 × 1013 VP/mL) |
| ELISA +
qPCR |
Antibody specificity to full and empty capsids,
used in tandem
with qPCR |
Expensive, not scalable, lacks accuracy and
precision, compounded
error |
| Analytical ultracentrifugation |
Separates capsid sedimentation rate of particles (buoyant densities) |
High accuracy, large volume of samples (300–400 μL), not scalable, time-consuming (6 h/sample) |
| LabChip electrophoresis
(Our method) |
Total protein/ssDNA ratio |
Scalable, fast turnaround time (6–15 min/sample), high throughput, requires low volumes (3–10 μL) and concentrations (>1 × 1011 GC/mL) |
