Size exclusion chromatography (SEC) separates molecules depending on differences in their size. SEC is frequently used as a polishing step during protein purification and can also be used for the analysis of protein homogeneity. In recent years the use of small-scale SEC for the analysis of low sample volumes or for the purification of small sample amounts has become more important; especially due to the development of suitable columns. Here, we performed a proof-of-concept study to the evaluate the performance of the AZURA® compact SEC system for the use of small-scale SEC applications and describe the system configuration for this application.
System configuration with 1/16" PEEK capillary connections from pressure sensor/pump outlet to UV detector inlet, red tubing 0.25 mm ID, blue tubing 0.13 mm ID
Chromatogram of gel filtration standard; (1) Thyroglobulin; (2) gamma-Globulin (3) Ovalbumin; (4) Myoglobulin; (5) Vitamin B12
Systemic pressure versus flow rate of the AZURA compact system (blue) pressure with capillary connections as described in Figure 1; red with additional 0.13 mm ID tubing to fraction collector
|Substances||Thyroglobulin, gamma-globulin, ovalbumin, myoglobin, Vitamin B12|
|Key words||FPLC, SEC, gel filtration, size exclusion chromatography, protein purification, protein separation, antibody purification|
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