Comparative study of Zorbax Bio Series GF 250 and GF 450 and TSK-Gel 3000 SW and SWXL columns in size-exclusion chromatography of proteins.

A reduction of the mean particle diameter of silica-based packings in the size-exclusion chromatography (SEC) of proteins to about 5 micron generates the expected increase in column plate number over the traditional 10 micron SEC columns, as demonstrated for the Zorbax Bio Series GF 250 and GF 450 and TSK-Gel 3000 SWXL columns. The slightly lower column efficiency of the TSK-Gel 3000 SWXL compared with the GF 250 column is compensated by the fact that the phase ratio of the 3000 SWXL column is higher by a factor of two. Hence both columns show nearly the same peak capacity of about 20-30. When the ionic strength of the eluent was changed by varying the salt concentration, the elution volume of proteins was found to be affected differently on the two columns. These alterations are attributed to the differences in the surface compositions of the two types of packings.