Fractionation of serum

Fractionation of Serum and Column Capacity. A Fractionation of human serum on the SW-3000 column is shown in Figure 3. 

Since antibodies occur in the globulin fraction of serum one predicts that the investigation of the fine structure of serum mucoproteins will become a problem of urgent immunological significance. It is doubtful however whether the carbohydrate group of mucoproteins can influence their immunological specificity.1 ... 

AlO. Aull, J. C., and McCord, W. M., Effects of posture and activity on the major fractions of serum pmtein. Amer. J. Clin. Pathol. 27, 52-55 (1957). 

Cemm will be determined by performing the described cholesterol assay directly on serum. CHDL will be determined on a separated, soluble HDL fraction of serum. Very low-density lipoproteins and low-density lipoproteins are selectively removed from serum by precipitation with magnesium-phos-photungstate reagent. 

H7. Henry, R. J., Golub, O. J., and Sobel, C., Some of the variables involved in the fractionation of serum proteins by paper electrophoresis. Clin. Chem. 3, 49 (1957). 

Robinson, J. C., B. S. Blumberg, and J. E. Pierce Studies on inherited variants of blood proteins. I. Two dimensional electrophoretic analysis of chromatographic fractions of serum proteins. J. Lab. Clin. Med. 60, 468 (1962). 

Anti-glucosyluronate-rhamnose antibodies were isolated from the globulin fraction of serum devoid of the anti-GlcA antibodies. The latter were... 

Ricketts, W.E., Kirsner, J.B., Palmer, W.L., Sterling, K. Observations on the diagnostic value of liver biopsy, tests of hepatic function, and electrophoretic fractionation of serum proteins in asymptomatic portal cirrhosis. J. Lab. Clin. Med. 1950 35 403-407... 

Johnson KL, Mason CJ, Muddiman DC, Eckel JE (2004) Analysis of the low molecular weight fraction of serum by LC-dual ESI-FT-ICR mass spectrometry Precision of retention time, mass, and ion abundance. Anal Chem 76 5097-5103. 

With a specific antibody, an analyte can often be quantified even in the presence of large amounts of extraneous materials that can obscure other detection systems (e.g., absorption or fluorescence spectra). Antibodies are glycoproteins found in the globulin fractions of serum and in tissue fluids. They are produced by vertebrates in response to the presence of an antigen, i.e., a substance that is recognized by the host to be foreign. Antibodies show a remarkable ability to bind selectively the antigen that stimulated their production. Their specificity may be... 

Since the usual ion-exchange resins have very limited use for chromatography of proteins, Peterson and Sober investigated adsorbents derived from cellulose, of which the most useful is DEAE-cellulose. It is prepared by treating a high-purity wood cellulose with 2-chloroethyldiethylamine hydrochloride to give a diethyl-aminoethyl (DEAE) derivative. This material has been used successfully for the fractionation of serum proteins and oligonucleotides. ... 

Serum total T4 concentrations were initially determined indirectly, using methods that measured the amount of iodine in a protein precipitate of serum (protein-bound iodine, PBI). In addition to hormonal iodine, the PBI tests also measured iodoproteins, iodotyrosines, inorganic iodine, and thyroglobulin. More specific T4 procedures involved the measurement of hormonal iodine in either a butanol extract of a serum protein precipitate (butanol-extractable iodine) or in a purified fraction of serum (I4 by column). These methods were useful because the iodine in T4 normally accounts for 80% to 90% of all iodine in serum. Both... 

Significant advances have been made during the last decade in the development of new techniques of paper, starch-, and agar-gel electrophoresis. Soon after the introduction by Smithies in 1955 of the technique of starch-gel electrophoresis as an important analytical tool for the fractionation of serum proteins (P21, R8), this procedure proved val-... 

Also, Hodson et al. (H17) applied Smithies technique for the characterization of alkaline phosphatases from different tissues (intestine, liver, bone, etc.). In their report they questioned the contention that the osseous system is the main contributing source of human plasma alkaline phosphatase (H17), and preferred liver alkaline phosphatase as the main fraction of serum alkaline phosphatase. Hodson et al. (H17) observed the occasional presence of alkaline phosphatase at the intestinal region also in sera after electrophoresis. 

K5. Keiding, N. R., Differentiation into three fractions of serum alkaline phosphatase and the behaviour of the fractions in diseases of bone and liver. Scand. J. Clin. Lab. Invest. 11, 106-112 (1959). 

A small but important fraction of serum copper is not bound to ceruloplasmin. Most of this non-ceruloplasmin-bound copper is attached to serum albumin. The amount of this fraction of serum copper varies in physiological and especially in pathological conditions. It can be measured since it will react with diethyldithiocarbamate directly, whereas ceruloplasmin-bound copper will react only after acidification. A method for the determination of direct reacting copper in serum has been published by Gubler et al. (G12). Unfortunately, this method is not very accurate, especially in the case of normal subjects where the figures are very low. More exact measurements can be obtained when the values are higher as in certain pathological states. 

Moreover, Neumann and Sass-Kortsak (N3) have shown recently that, in equilibrium with albumin-bound copper, there is a third fraction of serum copper bound to amino acids. This fraction is rather small compared even to the albmnin-bound fraction, but by virtue of its low molecular size it may have an important function in the transport of copper through membranes. Of the 21 amino acids present in human blood, histidine was the strongest binder of copper in competition with albumin. Some of the... 

The total serum copper levels are also usually reduced in patients with Wilson s disease (Bl, C4). Considering the low ceruloplasmin levels, this is not surprising. While the amounts of copper bound to ceruloplasmin are reduced, the non-ceruloplasmin-bound, direct-reacting fraction of serum copper is elevated in these patients (C5). Those with normal levels of ceruloplasmin have a high serum copper, since they have an elevated blood level of non-ceruloplasmin-bound copper together with normal amoimts of ceruloplasmin-bound copper (S9). 

It has long been known that the addition of organic solvents to aqueous solutions of proteins frequently causes their precipitation (Ml8). Among the first to achieve fractionation of serum protein with alcohol at low temperature were Liu and Wu (L13). Under controlled conditions, methanol can be made to precipitate all the globulins and leave the albumin in solution. 

Fractionation of serum proteins was achieved by affinity chromatography three main selenium fractions corresponded to SeP, albumin and GPx. Mass balance performed under different experimental conditions showed quantitative selenium recovery. The distribution of selenium between plasma GPx ( 20%), SeP ( 55%) and albumin ( 20%) was similar in both populations... 

Proper assessment of total serum calcium concentrations includes measurement of serum albumin concentrations. Hypoalbu-minemia, which may be associated with many chronic disease states, is probably the most common cause of laboratory hypocalcemia. Patients remain asymptomatic because the ionized fraction of serum calcium remains normal (reference range 4.4 to 5.4 mg/dL). A... 

H13. Hjalmarsson, S.-G., Preparative isotachophoresis—the effect of using ampholine of different pH ranges as spacer ions in the fractionation of serum proteins. Sci. Tools 22, 35-38 (1975). 

Lloyd E. 1968. Relative binding of strontium and calcium in protein and non-protein fractions of serum in the rabbit. Nature 217 355-356. 

The realization that the living organism is made up of a great number of individual proteins with different properties owed much to the discovery that they could be fractionated by taking advantage of their varying solubility in salt solutions of different concentrations. The pioneer here was the French biochemist Prosper Sylvain Denis (1799-1863) who in the 1850s published his results with salt fractionation of serum proteins. Salt fractionation became one of the most widely used methods for the separation of proteins and it remains a standard procedure to this day. 

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