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Process Media - Size Exclusion FAQ's

 

The following are Size Exclusion Process Media FAQ's. Please click on the corresponding tab below for the questions and answers.

  1. How do I know which size exclusion resin to use for my protein?

  2. Why is Size Exclusion Chromatography not used in large scale processes?

  3. Since Size Exclusion is not used in large scale process what alternatives are availble?

  4. How fast can I run an SEC column? If I run at a lower flow rate, do I get better resolution?


 

1.  How do I know which size exclusion resin to use for my protein?

Select a resin where the target protein is located in the middle of the separating range.  This will allow larger molecular weight as well as lower molecular weight special to be well resolved from the target. Please see our Solutions section for additional guidance in selecting your size exclusion resin.

 

2. Why is Size Exclusion Chromatography not used in large scale processes?

Size Exclusion Chromatography is a non-adsorptive mode.  This means that there is no calculated binding capacity and the amount charged to an SEC column is dependent on the column size, the concentration of the injection, the volume of the injection and the operating flow rate.  Typically the size of the column used when scaling up a process requires signicant increases in column size and height which are not economically feasible.

 

3. Since Size Exclusion is not used in large scale process what alternatives are availble?

Tosoh Bioscience often recommends using HIC as a attractive alternative to SEC.  Although HIC does not have the binding capacity of IEC, it is quite powerful at separating isoforms of the protein (such as aggregates and misfolded species) that may not be resolvable on ion exchange resins.

 

4. How fast can I run an SEC column? If I run at a lower flow rate, do I get better resolution

TOYOPEARL SEC resins are rated like all other TOYOPEARL media to withstand up to 3 bar. However, running at lower flow rates maximizes sample exposure to pore volume allowing for better resolution between molecules.