The other proposed mechanism is Secondary Exclusion Mechanism. This mechanism states that when a sample containing a mixture of small and large molecules is applied to a gel filtration column, the small molecules diffuse rapidly into the pores of gel, whereas large molecules will find relatively few unoccupied pores and move further down the column till they find the unoccupied pores. This results in the enhancement of separation of small and large molecules.
Applications:
- Purification.
- Desalting.
- Protein-ligand binding studies.
- Protein folding studies.
- Concentration of sample.
- Copolymerisation studies.
Relative molecular mass determination
Quiz:
Explain how size exclusion chromatography may be used for protein folding/refolding studies.
Reading: Werner et al., (1994) Refolding proteins by gel filtration chromatography. FEBS Letters 345, 125-130.
http://www.ncbi.nlm.nih.gov/pubmed/8200443
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