Preparation, characterization and storage of human ferritin for use as a standard for the assay of serum ferritin

Authors


M. Worwood, Department of Haematology, Welsh National School of Medicine, Heath Park, Cardiff CF4 4XN Wales

Abstract

Summary. This paper describes studies on the source, preparation, characterization and storage of human ferritin for use as a standard for the immunoassay of serum ferritin. Ferritin was prepared from the liver or spleen by methods including either ultracentrifugation or cadmium sulphate crystallization. Preparations were characterized by polyacrylamide gel electrophoresis in dissociating and non-dissociating buffers, iso-electric focusing, analysis of amino acid composition and measurement of protein content. The protein content of solutions of liver or spleen ferritin may be determined by the method of Lowry with bovine serum albumin as standard. Lyophilization under carefully controlled conditions in buffer containing high concentrations of albumin provides a stable preparation of ferritin. Accelerated degradation and collaborative immunological studies of two lyophilized preparations of ferritin, one from liver and one from spleen, indicate that either is an acceptable reference material.

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