Serum, fractionation

Figure 19.9. Chromatogram for stepwise elution of bovine serum albumin on a Vistec diethyl aminoethyl cellulose ion-exchanger, using stepwise increases in sodium chloride concentration in the mobile phase to achieve selective desorption. Proteins 1, serum fraction not adsorbed by column (includes y-gobuhn) 2,3,...

The latex was cleaned by ion exchange and serum replacement, which gave the cleaned latex plus six serum fractions. The cleaned latex and the serum samples were analyzed by conductometric titration. Also, the amount of anionic emulsifier in the serum was determined by Fyamine 1622 colorimetric titration and thin-film chromatography, and the amount of nonionic emulsifier by iodine-iodide colorimetric titration and thin-film chromatography. 

A polyvinyl acetate latex prepared by semi-continuous polymerization at 55° using a polymethacrylic acid-nonylphenol-poly-ethoxylate phosphate ester emulsifier and sodium persulfate-sodium formaldehyde sulfoxylate initiator (23). The latex was cleaned by ion exchange and serum replacement using both Nuclepore and Pellicon membranes, and the cleaned latex and serum fractions were analyzed by conductometric titration. In addition, the dried films were extracted with water and organic solvents, and the extracts were analyzed by infrared spectroscopy and thermo-gravimetric analysis. 

Accurate Kjeldahl determinations both on total serum and on serum fractions obtained by salting out show, according to Rein (Rl), less spreading than clinical electrophoresis in its present form. This indicates that the accuracy and reproducibility of the electrophoretic results must be increased in order to obtain greater clinical significance. 

Schade and Caroline (116) found the serum transferrin primarily in human blood serum fraction IV—3,4 (according to Cohn s nomenclature). The transferrin was purified and concentrated in fraction IV—7 by further subfractionations (30, 125, 126). Crystallized transferrin was prepared by crystallization in low dielectric solvents under low temperatures at controlled ionic strength and pH (81, 86). 

Saliva4 Serum fraction VIC Ovomucoid" Antifreeze glycoprotein" ... 

Pedersen, K. O. Ultracentrifugal Studies on Scrum and Serum Fractions. 

B8. Beam, A. G., and Kunkel, H. G., Localization of Cu in serum fractions following oral administration An alteration in Wilson s disease. Proc. Soc. Exptl. Biol. Med. 86, 44-48 (1954). 

Fig- 2. Percent of total FAEEs in fractions obtained by density gradient ultracentrifugation. Pooled sera from 15 intoxicated emergency-room patients were subjected to density gradient ultracentrifugation and serum fractions were isolated. Lipids were extracted and FAEEs were isolated by thin-layer chromatography and quantitated by gas chromatography. (From Doyle 1994 with permission.)... 

H22. Heilbronn, E., The pS-activity curve of a serum fraction IV-6-3 studied with butyrylcholine at pS above 3. Acta Chem. Scand. 12, 1879-1880 (1958). 

Serum and serum proteins have diverse functions which are now reasonably well understood for the industrially important cell lines, and which can generally be substituted by non-protein alternatives. Mammalian cells typically require a source of fatty acids, which were historically supplied by serum. To supply these, serum-free media usually contain plasma lipoprotein fractions, free fatty acids complexed to serum albumin or fatty acid/phospholipid microemulsions [61]. A high-density lipoprotein serum-fraction in medium containing bovine serum albumin was used by Seamans et al. [62] to replace serum in cultures of a recombinant antibody-producing GS-NSO cell line. Further, they found that the serum-fraction could be replaced with a commercially available non-protein-aceous lipid emulsion and a pluronic F-68/cholesterol emulsion. This gave equivalent growth and productivity (100 mg L ). 

Insulin-like activity in serum fractions was originally termed non-suppressible insulin-like activity, somatomedins (Daughaday et al., 1972) and multiplication-stimulating activity (Moses et al.,... 

For pediatric or micro serum Zn, serum was diluted 100-fold and a 10-//L sample was deposited in the furnace (Vieira and Hansen, 1981). The STPF technique was still too sensitive. The Zn line at 307.6 nm appears to have about 1000-fold less sensitivity which makes it too insensitive. It would be preferable to dilute the sample 200-fold and use a 5-//L aliquot. Because of the sensitivity, Zn has been determined in serum fractions separated by chromatography (Gardiner et al., 1981). Ten-//L urine samples were analyzed directly (Vieira and Hansen, 1981) in the furnace. 

C27. Coppa, G. V., Singh, J., Nichols, B. L., and Di Ferrante, N., Urinary excretion of disulfated disaccharides in Hunter syndrome Correction by infusion of a serum fraction. Anal. Lett. 6, 225-233 (1973). 

In order to understand which lipoproteins are involved in GGC trafficking and NKT cell activation, GGC was incubated with human serum in vitro, and various serum fractions, including the VLDL, LDL, and HDL fractions, were separated by fast protein liquid chromatography (FPLC). These fractions were then incubated with DCs to determine if they could activate NKT cells by measuring IFNy production from NKT cells. Interestingly, only the fraction corresponding to VLDL led to significant NKT cell activation whereas LDL and HDL fractions exhibited little or no NKT cell activation [61] (Fig. 7a,b). 

Improvements in the strategy of analyses have come from the realization that body fluids have structure and are made up of different physicochemical phases, which can be separated into subfractions for a more meaningful assay than that provided by a total fluid analysis. Thus, blood can be subdivided by centrifugation into cellular (erythrocytes, lymphocytes, and platelets) and noncellular (plasma or serum) fractions, while plasma or serum can be subdivided... 

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