1991 Volume 17 Issue 3 Pages 589-594
Endotoxin was separated from a macromolecular antibiotic SN-07 by ion exchange chromatography (IEC) and hydrophobic interaction chromatography (HIC). For IEC and HIC, endotoxin-free columns were prepared by washing the adsorbent gels with alkaline-ethanol followed by repeated wash with buffers of high and low concentrations of salt by turns.
Using an endotoxin-free column (Sepabeads FP-DA 13) for IEC, 95.5% of charged endotoxin was removed from the SN-07 fractions. For HIC, two types of adsorbent gel, Sepabeads FP-PH 12 and FP-PH 13, were used. FP-PH 13 has a higher ligand concentration than that of FP-PH 12. For both gels the differences in adsorption capacity for endotoxin at these salt concentrations of NaCl and (NH4) 2SO4 were slight. In HIC purification of SN-07, the removal of endotoxin involved in the SN-07 fractions after purification by IEC was as high as 92.8% for FP-PH 12 and 95.7% for FP-PH 13. This result corresponded to the adsorption capacity of both gels for endotoxin.
