Chromatography of Serum Proteins in Normal and Pathologic Sera: the Distribution of Protein-bound Carbohydrate

Chromatographic techniques have recently been introduced which provide new approaches to serum protein separation and characterization (1-4). Sober, Gutter, Wyckoff, and Peterson, using diethylaminoethyl (DEAE) cellulose, described the initial application of anion-exchange cellulose chromatography to the fractionation and identification of serum proteins (1, 2). Their study indicated the great potential usefulness of anion-exchange cellulose chromatography for the study of proteins, but considerable time and work was required to carry out a single determination. The system of analysis employed in these studies was adapted from a technique developed during a continuing investigation by Drs. Peterson and Sober (5) of chromatographic methods for protein separation. It differs from the technique described previously (2) principally in the use of a single gradient elution system and in modifications introduced in the present study for the purpose of decreasing the initial loss of euglobulins and reducing the time and work involved. These modifications made feasible the chromatographic examination of multiple individual serum samples and serum fractions. Serum samples of 20, 10, 2, and 1 ml. have been examined by this method. The present study reports the initial findings in the application of the modified chromatographic procedure to the examination of normal human serum and serum obtained in various diseases. The chromatographic fractions of normal serum were characterized by electrophoresis and by carbohydrate and cholesterol content. The chromatographic behavior of siderophilin (iron-binding protein, transferrin), vitamin B12-binding protein, thyroxin-binding protein, alkaline and acid phosphatases, myeloma protein and radioiodinated human serum albumin was also determined.