Isolation of individual protamines by high-performance liouid chromatography

Using as an example the protamine from the gonads of the sturgeon Acipenser stellatus, a procedure has been developed for isolating homogeneous protamines (stellins) by high-performance liquid chromatography which, as compared with the ion-exchange chromatography used for these purposes at the present time [i, 2], permits a considerable shortening of the time of isolation and the elimination of the procedure for desalting the proteins obtained, which frequently leads to their destruction. Chromatography was performed with the aid of a model 8800 liquid chromatograph (USA) fitted with a UV-spectrophotometric detector with a flowthrough cell (wavelength 220 nm; rate of flow 1.0 ml/min). Zorbax ODS column (4.6 × 250 nm) at a column temperature of 35°C. The proteins were eluted with aqueous ethanol, using a concentration gradient of ethanol of from 0 to 30%. To prevent the irreversible binding to the sorbent that is characteristic for basic proteins [3], trifluoroacetic acid was added to the mobile phase in a concentra