TSK-Toyopearl gels for the preparative separation of sterol carrier protein2 from rat liver

The application of TSK-Toyopearl gels to the preparative separation of a basic and low-molecular-weight protein, sterol carrier protein2 (SCP2), was studied. SCP2 was purified from 105,000 g supernatant of rat liver (S105) by ion-exchange chromatography on CM-Toyopearl 650M and gel permeation on Toyopearl HW60S. Separation of S105 by CM-Toyopearl 650M was carried out at a high flow-rate in the presence of 10% (v/v) glycerol, a stabilizer of the protein. Toyopearl HW60S showed a significant ion-exchange effect on the elution of SCP2. Using an elution buffer of ionic strength of 45 mM, a highly efficient purification of SCP2 on HW60S was achieved. SCP2 was purified approximately 5000-fold to apparent homogeneity with an overall yield of 69%.