Isolation of mouse transferrin using salting-out chromatography on sepharose CL-6B

Abstract

A new method for the isolation of considerable quantities of mouse transferrin is described. This technique employs salting-out chromatography on Sepharose CL-6B, a new step in the preparation of plasma proteins. This step is followed by ion-exchange chromatography on DEAE-Sepharose CL-6B and gel filtration on Sephacryl S-200. The isolated mouse transferrin was shown to be pure by immunoelectrophoresis, sodium dodecyl sulphate-polyacrylamide gel electrophoresis and by the 465 nm 410 nm ratio of absorbances being 1.41. The molecular weight was determined tobe about 77 500. The advantages of this procedure are that it is reproducible, gives a high recovery, and can be extended to a larger scale. The advantage over other protein purification techniques is its general utility, due to the fact that there is no need for species-specific antibodies. The application of this method offers a rapid purification of sufficient quantities of mouse transferrin essential for the elucidation of biological functions of this protein and investigations of its molecular structure.