Bio-Rad Nuvia™ cPrime™ Media User Manual

Page 13

Section 7

Sanitization and Regeneration

After each chromatography run, the packed media bed should

be washed to remove reversibly bound material and prepare the

column for the subsequent run. This cleaning process is achieved

by washing the column with 2–6 column volumes of 1–2 M

NaCl followed by 2–6 column volumes of 0.1 N NaOH to remove

remaining proteinaceous impurities. Washing should be conducted

until absorbance returns to baseline. The column is now ready to

be sanitized in 1.0 N NaOH.

After sanitization, to equilibrate the column we recommend

applying 4–6 column volumes of a solution such as 60 mM NaOAC

(sodium acetate)

Note: if the column no longer yields reproducible results, the

media may require additional cleaning to remove strongly bound

contaminants. Acceptable cleaning agents include 25% acetic

acid, 8 M urea, 1% Triton X-100, 6 M potassium thiocyanate,

70% ethanol, 30% isopropyl alcohol, 1 N NaOH, or 6 M guanidine

hydrochloride.