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Cracked columns: causes, prevention, and regeneration

Some qEV column users have reported cracks forming in the resin bed after one or two uses. This article explains why cracking occurs, how to prevent it, how to regenerate affected columns, and what is currently known about performance following regeneration.

Why do some columns crack after use?  

qEV columns contain a bed of porous resin beads that are sensitive to temperature changes. 

Rapid temperature shifts (temperature shock) can cause the resin bed to crack. 

Cracking most commonly occurs when a cold sample or buffer (e.g. 4 °C) is applied to a column that is at room temperature. 

How can I prevent columns from cracking?  

To minimise temperature shock to the resin bed: 

  • Equilibrate samples and buffers to room temperature before use.  
  • Ensure qEV columns are at room temperature before use. 
  • Ensure qEV columns are stored appropriately. We recommend storing qEV columns upright at room temperature. Some qEV users choose to use a storage solution which needs to be kept between +4 and +8 °C. If so, allow the column to reach room temperature before use. 

What do column cracks look like? 

Cracks may appear as: 

  • Small, dot-like features within the resin bed, or  
  • A visible separation between the resin and the column wall.  

 Do qEV column cracks impact performance? 

Cracks represent a potential source of variation and may influence the elution profile. 

For this reason, we recommend regenerating cracked columns before further use. 

How to regenerate cracked columns  

Cracks can typically be resolved by placing the column in a refrigerator (+4 to +8 °C). 

In our experience, cracks usually clear within one hour. 

Once cracks are no longer visible, the column can be returned to room temperature before reuse. 

 What do we know about the performance of regenerated columns?  

Based on in-house testing: 

  • No difference in particle size or concentration was observed between regenerated and unaffected qEVoriginal columns (35 nm and 70 nm), measured by TRPS. Particles sized between 70 and 500 nm in bovine plasma were analysed. 
  • Regenerated columns showed consistent elution profiles and particle measurements for up to two subsequent uses when tested with carboxylated polystyrene particles (70–1000 nm). 

Based on the data currently available, our results suggest that regenerated columns can be reused without concern for performance. 

What don’t we know yet?  

At this time: 

  • Protein elution profiles have not been assessed. 
  • EV biomarkers and other particle types (e.g. lipoproteins) have not been evaluated following column regeneration.