HomeProtein PurificationSeparation Options with Superdex®

Separation Options with Superdex®

Superdex® is produced with different average particle sizes and different selectivities (see Table 2.2).

  • Use the prepacked columns with Superdex® Peptide, Superdex® 75, and Superdex® 200 Increase for small-scale preparative purification and analytical runs.
  • Use columns prepacked with Superdex® 30 prep grade, Superdex® 75 prep grade, or Superdex® 200 prep grade media for preparative applications.
  • Use bulk media of Superdex® 30 prep grade, Superdex® 75 prep grade, and Superdex® 200 prep grade for preparative purification with larger sample volumes of up to 13 mL.
  • 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, Chapter 1).
  • Start with Superdex® 200 Increase when the molecular weight of the protein of interest is unknown. Superdex® 200 Increase or Superdex® 200 prep grade are especially suitable for the separation of monoclonal antibodies from dimers and from contaminants of lower molecular weight, for example albumin and transferrin.
  • Start with Superdex® Peptide or Superdex® 30 prep grade for separations of peptides, oligonucleotides and small proteins below Mr 10 000.
  • Exposure to temperatures outside the range 4 °C to 40 °C will negatively affect the efficiency of the packed bed and the column will need to be repacked.
Table 2.2.Separation options with Superdex® media

* HiLoad is packed with Superdex® prep grade.
† For maximum resolution, apply as 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. § prep grade.
** Typical pressure drop over the packed bed.
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