Chromatographic Scale-Up Procedures

T able 14. Effects of Process Parameters on Resolution and Throughputs ll 

Parameter Resolution varies with Throughput varies with 

The scaleup considerations of column chromatography for protein isolation has been described by Charm and Matteo.l l When several hundred liters of a protein feedstream must be treated, the resin may be suspended in the solution, removed after equilibration by filtration and loaded into a column from which the desired proteins may be eluted. Adsorption onto a previously packed column was not recommended by them since they feared suspended particles would clog the interstices of the column, causing reduced flow rates and increased pressure drops across the column. The reduced flow rate may lead to loss of enzyme activity because of the increased time the protein is adsorbed on the resin. 

It is important that all of the resin slurry be added to the column in one operation to obtain uniform packing and to avoid the formation of air pockets in the column. An acceptable alternative, described by Whatman,I allows the addition of the adsorbent slurry in increments. When the resin has settled to a packed bed of approximately 5 cm, the outlet is opened. The next increment of the slurry is added after the liquid level in the column has dropped. It is important that the suspended adsorbent particles do not completely settle between each addition. 

LadischI J has worked with a variety of column sizes ranging from 2 to 16 mm in diameter and 10 to 600 cm in length. His experience is that published semi-empirical scaleup correlations are useful in obtaining a first estimate on large scale column performance. 

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