Column chromatography with Sepharose beads containing

Column Chromatography. Sepharose beads containing covalently linked gangliosides (0.2 ml packed volume) were placed into a pasteur pipette containing a small amount of glass wool. Columns were washed with HEM containing 50 ug/ml bovine serum albumin (3 ml). Interferon solutions in MEM-albumin (1 ml) were placed on the columns, which were eluted with MEM-albumin at a flow rate of no more than one drop per minute. Fractions of 1 ml were collected and interferon titers determined in each fraction after serial two-fold dilution. Columns onto which mouse fibroblast interferon had been loaded, were eluted with MEM-albumin first, then with 0.07 M N-acetylneuraminyl lactose at pH 2. 

In this method, proteins of interest are purified by use of epitope tag. The epitope tags consist of small stretch of amino acids that are fused to proteins of interest. The proteins with such epitopes are purified via column chromatography by passing an extract of proteins over a matrix such as Sepharose beads (Sigma Aldrich Co., St. Louis, MO) containing a protein with affininty for the epitope. During such column chromatography, the... 

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