CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. XL. QUANTITATIVE SEPARATION AND DETERMINATION OF THE PROTEIN COMPONENTS IN SMALL AMOUNTS OF NORMAL HUMAN PLASMA 1

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Abstract
The procedure of Cohn et al.(Jour. Amer. Chem. Soc/72(1): 465-474. 1950.) was applied to the analysis of 5 ml. samples of the blood plasma of normal men. Apparatus comprising sintered glass filter funnels immersed in a cold bath at -5[degree]C and connected to a regulated vacuum source was employed for the separation of the plasma proteins. Stock buffer soln., when em-ployed in the prescribed vols., yielded the desired conditions without further adjustment. Four fractions were separated, containing principally (1) serum albumins, [alpha]1-lipoproteins, [beta]1-metal combining protein, various [alpha]2-glycoproteins in Fraction IV+V; (2) an [alpha]1-glycoprotein and other trace components in Fraction VI; (3) [gamma] -globulins in Fraction II; (4) [beta]1-lipoproteins, caeruloplasmin, isohemagglutinins, and the major components of the clotting process in Fraction I+III. The avg. distr. of total protein among the fractions was 65.6, 2.2, 10.8 and 22.0%, respectively. Cholesterol and phospholipids were concentrated nearly quantitatively with the lipoproteins in Fractions I+III and IV+V, whereas protein-bound carbohydrate was distr. more evenly. Concns. of serum albumins, [alpha]- and [beta]-lipoproteins and [gamma]-globulins were detd. Electrophoretic distrs. in the fractions were reported. Added amts. of various pure proteins were in each case recovered in the appropriate fraction. The separated protein components were in an unaltered state, and retained unimpaired biological activity.