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SEPARATION AND CHARACTERIZATION OF HUMAN SERUM

During the early phase of transport of exogenous fats, lymph and blood assume a milky appearance due to an increase of large fat particles, known as chylomicrons. Separation can be achieved by chemical means; protamine sulfate (1), toluidine blue (2), heparin and calcium chloride (3, 5), polyvinylpyrrolidone (4); however, ultracentrifugation (6) is the method commonly used. Chemical characterization of lymph and serum chylomicrons has not been satisfactorily achieved; this may be owing to: (a) variation of laboratory procedures, which do not yield a product of constant composition; (b) variation in source: human lymph (7-14), human serum (12), rat lymph (2, 15, 16), dog lymph (17, 18), and dog plasma (19); (c) limited number of reports on chylomicron composition and lack of comparative studies between chylomicrons derived from lymph and blood. Another problem is the relationship between chylomicrons and serum lipoproteins. Robinson (15) found no protein in chylomicrons separated from rat chyle; other reports (2, 9, 11, 14, 16, 17) however, show small percentages present in human and rat chyle chylomicrons. The nature of this protein is unknown. Middleton (20) reported the presence in human serum chylomicrons, of a protein antigenically related to serum beta lipoproteins. Recently, Robdell (21) found that serine, threonine, and aspartic acid are the N-terminals of the chylomicron protein from human lymph serum. Our study concerns: (a) Separation, purification, and chemical composition of chylomicrons from serum of people in absorptive phase; (b) separation and characterization of their protein moiety; (c) participation of the chylomicron protein in a clearing factor system "in vitro"; (d) comparison of the chylomicrons with ultracentrifugally separated creamy top fractions from preincubated mixtures of serum and coconut oil.