AAV – purity and integrity
Monitoring each step in the purification of AAV
Undesired AAV sample characteristics to consider when optimizing the downstream process
- Failure to remove host-cell debris or residual DNA
- Loss of particle morphology or integrity
ALl these types of contaminants can easily be identified and quantified by using the automated image analysis software tools in MiniTEM
After 1st purification step After 2nd purification step After final purification step
Types of contaminants that may appear
Large size impurities Small size impurities
Aggregates Residual DNA
Automated MiniTEM analysis of purity and level of debris
MiniTEM automatically images, detects and classifies particles and debris. Since automation enables detection and analysis of a far larger number of particles than is feasible with manual operation, the results are more consistent, statistically significant and reproducible.
Undetected particles Detected primary particles and debris
Quantified particles Time saving through automated MIniTEM analysis
Capsid integrity analysis with MiniTEM
The proportion of broken AAV capsids in a sample can be measured using MiniTEM as negative stained samples of AAV particles display variations in staining patterns correlated to capsid integrity status.
Intact AAV particles appear as lacking internal staining as opposed to broken particles that exhibit internal dark staining due to disrupted virus capsid structures.