Detoxification by gel filtration

Equilibrate the column with one bed volume growth medium without serum. 

Remove the cell-free medium from the culture, leaving a minimum amount of medium for the cells to survive for approximately 2h. If the cells are not adherent, or immobilized by other means, they must be removed by, for example, centrifugation. 

Apply the cell-free medium to the column and start the recorder and fraction collector. 

Elute growth medium without serum (one bed volume) and collect suitable fractions (e.g. 1/10 to 1/5 of the volume applied to the column). 

Pool the fractions containing the proteins (peak adsorption at 280 mn) to give a total volume equal to the volume applied to the column. 

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Procedures are given for two different methods detoxification by dialysis and detoxification by gel filtration. 

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