The chromatographic purification of lysozyme
from the CEW solution using cation-exchange cryogel
were carried out at a constant flow velocity of 2 cm·min−1,
according to our previous work [10], using 20 mmol·L−1
sodium phosphate buffer (pH 7.8) as a running buffer.
The column was equilibrated with running buffer.
Thereafter, a feedstock of 27.2 ml was loaded and the
column was washed with the same buffer.
One-step and sequential elution were performed
with 0.5 mol·L−1 NaCl and various concentrations of
NaCl, respectively, in running buffer. The chromatography
processes were monitored using an on-line flowthrough
UV spectrometer at 280 nm as reported in our
previous work [12]. The column effluent was recorded
over time and collected by 1.6 or 0.8 ml of each aliquot
for further analysis. Finally, column cleanings
were performed with 0.1 mol·L−1 HCl, 1.5 mol·L−1
NaCl and deionized water.