advantages and disadvantages of gel filtration chromatography 7 | Page 4

growth. Cleaning the column thoroughly and keep it under cold
conditions in case that the column will not be used for a long time.
3. Elution earlier than expected
Reason: Ionic aggregates and complexes due to carbohydrate chains
(glycoproteins).
Recommended solution: Include 150mM NaCl or up to 10% betaine in
buffer.
Reason: Complexes associated via SH-groups.
Recommended solution: Add reducing agent (e.g. DTT) to buffer.
Reason: Hydrophobic aggregates.
Recommended solution: Add 10-20% ethanol or other organic solvents to
buffer.
Reason: Gel filtration of micelles/protein aggregates.
Recommended solution: Decrease buffer detergent concentration until
below CMC or use a different detergent (stay below its CMC).
Remember to check that your sample and your column are stable in the
buffer before using special additives to aid solubilisation.