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AAV – purity and integrity

Monitoring each step in the purification of AAV

Undesired AAV sample characteristics to consider when optimizing the downstream process

  • Aggregation
  • 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. 

 

 

 

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