Kidney purification

Yokomizo, T., T. Izumi et al. 1993. Enzymatic inactivation of leukotriene B4 by a novel enzyme found in the porcine kidney. Purification and properties of leukotriene B4 12-hydroxydehydrogenase./Bio/ C/iem268(24) 18128-18135. 

Cellulose acetate films, specially cast to have a dense surface and a porous substmcture, are used in reverse osmosis to purify brackish water (138—141) in hollow fibers for purification of blood (artificial kidney) (142), and for purifying fmit juices (143,144) (see Membrane technology). 

Another application shows the preparative purification and polishing of a therapeutic fusion protein with a humanized recombinant IgG protein. The fusion protein was expressed by the fermentation of baby hamster kidney cells. The filtered culture supernatant (155 liters) contained 2.2 g of IgG and 75.5 g of total protein. After the immunoglobulins were isolated by expanded bed adsorption and rebuffering, the IgG fraction was bound to Fractogel EMD SOj (M). This column achieved baseline separation of complete antibodies (fusion protein) from small amounts of antibodies lacking the fusion part. The resulting highly purified IgG fraction (110 ml) was diluted to 150 ml and... 

The procedure for purification of Na,K-ATPase in membrane-bound form from the outer renal medulla of mammalian kidney offers the opportunity of exploring the structure of the Na,K-pump proteins in their native membrane environment. The protein remains embedded in the membrane bilayer throughout the purification procedure thus maintaining the asymmetric orientation of the protein in the baso-lateral membrane of the kidney cell in the purified preparation. This preparation has been particularly useful in studies of ultrastructure, protein conformation and for... 

Kikugawa, K., Beppu, M. and Sato, A. (1995). Extraction and purification of yellow-fluorescent lipofuscin in rat kidney. Gerontology 4/(Suppl. 2), 1-14. 

Bouizar, Z., Fouchereau-Person, M., Taboulet, J., Moukhtar, M.S., and Milhaud, G. (1986) Purification and characterization of calcitonin receptors in rat kidney membranes by covalent cross-linking techniques. Eur. J. Biochem. 155, 141-147. 

After its purification, sterile filtration and aseptic filling, human urokinase is normally freeze-dried. Because of its heat stability, the final product may also be heated to 60 °C for up to 10 h in an effort to inactivate any undetected viral particles present. The product utilized clinically contains both molecular mass forms, with the higher molecular mass moiety predominating. Urokinase can also be produced by techniques of animal cell culture utilizing human kidney cells or by recombinant DNA technology. 

CIEF was also used to follow the production of recombinant antithrombin III (r-AT Iff) in cultures of hamster kidney cells.111 r-AT III inhibits serine proteases such as blood factors (IXa, Xa, and XIa) and thrombin. Interference by the media from which the samples were collected posed some difficulties because some of the media components have similar characteristics to those of the compounds of interest. CIEF was used to determine the pis of the separated components after sample purification by HPLC. Three major peaks showed pis of 4.7, 4.75, and 4.85, and three minor peaks had pis of 5.0, 5.1, and 5.3. These data closely resembled the data already published for serum AT III based on conventional IEF. 

Haemodialysis equipment parts, membranes for artificial kidneys, blood purification. .. 

Selected entries from Methods in Enzymology [vol, page(s)] Detergent-resistant phospholipase Ai from Escherichia coll membranes, 197, 309 phospholipase Ai activity of guinea pig pancreatic lipase, 197, 316 purification of rat kidney lysosomal phospholipase Ai, 197, 325 purification and substrate specificity of rat hepatic lipase, 197, 331 human postheparin plasma lipoprotein lipase and hepatic triglyceride lipase, 197, 339 phospholipase activity of milk lipoprotein lipase, 197, 345. 

J. J. Marshall and C. M. Lauda, Purification and properties of phaseolamin, an inhibitor of a-amylase, from the kidney bean, Phaseolus vulgaris, J. Biol. Chem., 250 (1975) 8030-8037. 

An amidrazone (58) derived from 5-amino-5-deoxy-L-fuconolactam was found to inhibit a recombinant human a-L-fucosidase with a K -value of 820 nmol/1 [ 111 ]. A simple synthesis of 1,5-dideoxy-1,5-imino-D-arabinitol (59), previously prepared by Ganem et al. [49] as a potential maimosidase inhibitor, was applied to the affinity purification of a-L-fucosidase from bovine kidney by an improved method and the characterization of the enzyme thus obtained [112]. The relatively low affinity of this compound to the enzyme (Kj 2.2 pmol/1 at pH 7) compared to 1-deoxyfuconoJirimycin (51) turned out to be advantageous in terms of enzyme recovery and yield. Structurally related, suitably protected 5-amino-5-deoxy-D-arabinopyranose (60), was coupled with a N-acetyl-6-deoxy-6-thio-D-glucosaminide (61) to give a stable thioglycoside (62) [113]. 

Kirkbride (1987) described the estimation of diazinon in human omental tissue (fatty tissue) after a fatal poisoning. In this method, the tissue was pulverized and extracted with acetone. After extract concentration and purification by sweep co-distillation and Florisil fractionation, diazinon was measured by gas chromatography (GC) with nitrogen-phosphorus detection (NPD). After another fatal diazinon poisoning, diazinon was quantified by GC/electron capture detection (ECD) and GC/flame ionization detection (FID) by Poklis et al. (1980). The diazinon in human adipose, bile, blood, brain, stomach contents, kidney, and liver was recovered by macerating the sample with acetonitrile followed by the addition of aqueous sodium sulfate and extraction into hexane. Following an adsorption chromatography clean-up, the sample was analyzed. 

Odessey, R. Purification of rat kidney branched-chain oxo acid dehydrogenase complex with endogenous kinase activity. Biochem. J., 204, 353-356 (1982)... 

Lawson, R. Cook, K.G. Yeaman, S.J. Rapid purification of bovine kidney branched-chain 2-oxoacid dehydrogenase complex containing endogenous kinase activity. FEBS Lett., 157, 54-58 (1982)... 

Lee, H.Y. Hall, T.B. Kee, S.M. Tung, H.Y.L. Reed, L.J. Purification and properties of branched-chain a-keto acid dehydrogenase kinase from bovine kidney. BioFactors, 3, 109-112 (1991)... 

The spleen is an organ, the main functions of which are the formation and purification ofblood. Blood from the spleen and intestine is passed through the portal vein to the liver for further reactions. The functions of the lungs, kidneys, and liver are described later in the chapter. The coronary arteries, which branch from the aorta, supply blood to the muscles of the heart. 

Liquid chromatography cleanup on a LiChrosorb Diol column has been further proposed for the offline purification of chloramphenicol residues from bovine muscle and eggs (32). An online approach based on reversed-phase principles has also been described for isolation of chloramphenicol residues from swine kidney by an automated column switching system (63). Use of a protein exclusion column (Hisep) has been also suggested in an online trace-enrichment method for the determination of chloramphenicol in animal tissues (52). By employing a column-switching system, all chloramphenicol that eluted from the protein exclusion column was trapped at the entry of a 5 m Supelcosil LC-18 preconcentration column, to be subsequently back-flashed into the analytical column. 

Thus, the main impurities are water, HCOOH and (CH3)2NH. Small amounts of HCN, formed by photolysis, and CO, formed by thermal decomposition, are also present. In the purification of a commercial product, it is kept with molecular sieves (4A or 5A) for 1-4 days to dehydrate. Then it is shaken with BaO for 1-2 days and the supernatant is distilled twice at 20 mmHg under nitrogen. All these operations must be carried out in the absence of light. The distillate should be stored under nitrogen gas and used as soon as possible. DMF has toxic effects, particularly on the liver and kidney and care should be taken in its handling. 

Purification of the enzyme from kidney has been reported for hog (68, 71, 74), beef (72), sheep, lamb, rabbit (70) and human (76). 

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