Separation Options with Superose®

Extracted from Size Exclusion Chromatography Principles and Methods, (PDF)
GE Healthcare, 2014

Superose® is produced in different average particle sizes and with two different selectivities (Table 3.1). Superose® 6 and 12, as well as Superose® 6 Increase prepacked in 3.2/300 columns are an excellent choice when working with very small sample volumes and for high resolution small-scale separations. Superose® 6 Increase 5/150 GL is the first choice for rapid screening and for applications where buffer consumption needs to be low. The column is also an excellent choice for for purity checks. Superose® 6 and 12 as well as Superose® 6 Increase 10/300 GL are designed for highresolution preparative-scale purification, protein analysis, and protein characterization.

Use the prepacked columns with Superose® 6 Increase and Superose® 12 for small-scale preparative purification and analytical runs.

Use bulk media of Superose® 6 prep grade and Superose® 12 prep grade for preparative purification.

 

Table 3.1. Separation options with Superose® media

Product Fractionation range, Mr (globular proteins) Sample loading capacity* Maximum pressure drop over the packed bed Recommended operational flow rate
Superose® 6 Increase 10/300 GL 5 × 103 to 5 × 106 25 to 500 μl 3.0 Mpa, 30 bar, 435 psi 0.5 ml/min
Superose® 6 Increase 5/150 GL 5 × 103 to 5 × 106 4 to 50 μ 3.0 Mpa, 30 bar, 435 psi 0.3 ml/min
Superose® 6 Increase 3.2/300 5 × 103 to 5 × 106 4 to 50 μl 3.0 Mpa, 30 bar, 435 psi 0.04 ml/min
Superose® 6 10/300 GL 5 × 103 to 5 × 106 25 to 500 μl 1.5 MPa, 15 bar, 217 psi 0.1 to 0.5 ml/min
Superose® 6 3.2/300 5 × 103 to 5 × 106 < 50 μl 1.2 MPa, 12 bar, 175 psi 0.04 ml/min
Superose® 6 prep grade
(Bulk medium)
5 × 103 to 5 × 106 0.5% to 4% of total column volume Column-dependent up to 40 cm/h
 
Superose® 12 10/300 GL 5 × 103 to 5 × 106 25 to 500 μl 3.0 MPa, 30 bar, 435 psi 0.5 ml/min
Superose® 12 3.2/300 5 × 103 to 5 × 106 < 50 μl 2.4 MPa, 24 bar, 350 psi 0.04 ml/min
Superose® 12 prep grade
(Bulk medium)
5 × 103 to 5 × 106 0.5% to 4% of total column volume Column-dependent up to 40 cm/h

* For maximum resolution, apply as a small sample volume as possible. Note that sample volumes less than 0.5% normally do not improve resolution.

See Appendix 5 to convert linear flow (cm/h) to volumetric flow rates (ml/min) and vice versa.
Typical pressure drop over the packed bed.

The maximum pressure drop over the packed bed is a typical value. Note that the actual value is individual for each column and needs to be determined (see Setting column pressure limits).

Materials

     
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