How should a column be cleaned if it is typically used to analyze protein samples?



Last Updated: 4/23/2018

A fairly extensive column cleaning procedure may be necessary to completely remove
protein contaminants because of the complex nature of protein samples. At each
cleaning step, one third to one half of the normal mobile phase flow is recommended
to prevent over pressurizing the column. If strong ionic interactions between proteins
and the stationary phase are suspected then start cleaning with a denaturant such as
6 M guanidine hydrochloride or 10 % DMSO. If the protein is relatively hydrophobic,
start by flushing out buffer with 95-100 % water, then clean out the hydrophobic
proteins with a gradient from 95 % water/5 % acetonitrile up to 5 % water/95 %
acetonitrile over 3-5 column volumes. If pressure is abnormally high and column frit
contamination is suspected, the column can then be reverse-flushed. After using guanidine,
the column should be washed with 40-50 column volumes of water or buffer to
adequately flush the column. However, there is no guarantee that the proteinaceous
material will be completely removed. For more information, please refer to the bioZen
column care notes.