Fractionation of Urine to Allow Desmosine Analysis by Radioimmunoassay

1992 ◽  
Vol 29 (1) ◽  
pp. 72-78 ◽  
Author(s):  
Barry Starcher ◽  
Marti Scott
Keyword(s):  
Ion Exchange ◽  
Affinity Chromatography ◽  
Human Urine ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
The Other ◽  
Normal Adult ◽  
Average Adult

The present study was designed to re-evaluate the radioimmunoassay for desmosine in urine, which is currently used as a measure of elastin metabolism. Using ion exchange chromatography, gel filtration and affinity chromatography it was shown that at least five other compounds in hydrolysates of human urine competed for desmosine in the RIA. Fractionating the urine prior to hydrolysis with acetone removed one of the major contaminants. The other contaminants could subsequently be removed by extracting the urine hydrolysate with a mixture of chloroform/ethanol (60:40). Samples from nine normal adult urines showed that an average of 45% of the RIA competing material in unfractionated urine was not desmosine. The final extracted residue retained all of the desmosine and only 16% of the original solids. The average adult urine contains approximately 50 pmol desmosine/mg creatinine, reflecting a daily turnover of between 3 and 4 mg of elastin per day.

Evaluation of dialysis treatment in uremic patients by gel filtration of serum

1990 ◽  
Vol 36 (11) ◽  
pp. 1906-1910 ◽  
Author(s):  
J Osada ◽  
T Gea ◽  
C Sanz ◽  
I Millan ◽  
J Botella
Keyword(s):  
Ion Exchange ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
The Other ◽  
Control Group ◽  
Dialysis Treatment ◽  
Additional Information ◽  
Molecular Masses ◽  
Two Peaks

Abstract A group of substances of molecular masses between 300 and 1500 Da have been found to be toxic metabolites in patients with uremia. We determined the concentration in serum of these molecules in the following groups of patients: two hemodialyzed groups (one with cuprophane and the other with polyacrylonitrile dialyzers), one group treated with continuous ambulatory peritoneal dialysis, one group of nondialyzed azotemic patients, and one control group of healthy persons. Ultrafiltrates of the subjects' sera were fractionated on Sephadex G-15 followed by ion-exchange chromatography. Eluates were monitored by absorbance at 254 and 206 nm. Partially characterized peaks P1 and P2, obtained by gel filtration, correlated with the concentration of creatinine in serum; their concentrations were significantly (P less than 0.01) larger in hemodialyzed groups than in peritoneal dialyzed or in nondialyzed azotemic patients. After ion-exchange chromatography, two peaks (P'5 and P'6) correlated with serum creatinine and also were larger in hemodialyzed patients than in the other groups. Apparently, adequate discrimination is obtained by gel-filtration analysis and further analysis by ion-exchange chromatography does not provide additional information in most of the affected patients.

scholarly journals Current Trends in Protein Purification : A Review

2020 ◽  
pp. 279-310
Author(s):  
Angela Boxi ◽  
Isha Parikh ◽  
Radhika B S ◽  
Shryli K S
Keyword(s):  
High Pressure ◽  
Liquid Chromatography ◽  
Ion Exchange ◽  
Affinity Chromatography ◽  
Protein Purification ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Current Trends ◽  
Comparative Information

The present review is based on papers published between 1990 and 2020 and gives Comparative information about the most common protein purification techniques Gel-Filtration, Chromatography, Ion-Exchange Chromatography, Electrophoresis, Affinity Chromatography, and Dialysis, High-Pressure Liquid Chromatography. and their applications.

scholarly journals Influence of various levels of L-asparaginase II purification on the cytotoxicity, DNA level, and apoptosis in Hep-2 cells

2010 ◽  
Vol 4 (2) ◽  
pp. 46-52
Author(s):  
Hayfa H. Hassani ◽  
Rawa'a J. Toma
Keyword(s):  
Ion Exchange ◽  
Cancer Cells ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
The Other ◽  
Proteus Vulgaris ◽  
Bacterial Enzyme ◽  
M Phase ◽  
Induced Apoptosis

The genetic effects of several concentrations of L–Asparaginase II (ASNase II), produced by Proteus vulgaris strain Pv.U92, at various levels of purification (ultrasonication, precipitation, ion-exchange chromatography and gel filtration chromatography) on cancer cells line of Hep–2 were studied. This bacterial enzyme with concentration 4 U/ml at gel filtration level was revealed a putative cytotoxicity against cancer cells in comparison with other concentrations and steps of purification were used in this work. Moreover, 4 U/ml of ASNase II at gel purification level has a distinguished role on arrest cancer cells division of Hep–2; it was reduced the content of DNA at each phase of cancer cell cycle particularly at G2/M phase, the level of DNA was 3%. On the other hand, the partial purified enzyme, L–ASNase II, was induced apoptosis by both levels of purification ion–exchange and gel filtration, the apoptotic fractionation was 0.86 and 0.7 respectively .

Studies on salivary and pancreatic lipases of the pre-ruminant calf

1982 ◽  
Vol 49 (3) ◽  
pp. 347-360 ◽  
Author(s):  
Joyce Toothill
Keyword(s):  
Ion Exchange ◽  
Affinity Chromatography ◽  
Pancreatic Lipase ◽  
Lipase Activity ◽  
Lipolytic Activity ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Maximum Activity ◽  
Exchange Chromatography ◽  
Single Enzyme

SUMMARYSalivary and pancreatic lipases of the pre-ruminant calf have been studied using ion-exchange chromatography and gel filtration. In addition, pancreatic lipase has been fractionated using concanavalin A-affinity chromatography. The effects of 5,5′-dithiobis(2-nitrobenzoic acid), organic solvents and trypsin on pancreatic lipase have also been investigated. The effect of taurodeoxycholate on the lipolytic activity of the 2 lipases has been compared. Salivary lipase behaved as a single enzyme on ion-exchange chromatography, and gel filtration gave a mol. wt value of 52000 for the enzyme. Although pancreatic lipase appeared to be a single enzyme on gel filtration, with a mol. wt of almost 80000, the lipase was shown by ion-exchange and affinity chromatography to consist of at least 2 enzymes of mol. wts 72000 and 60000, and did not require colipase for maximum activity in the presence of high concentrations of bile salts. Colipase-dependent lipase, mol. wt about 45000, and probably amounting to not more than 10% of the total activity, was also present. This was the predominant form only after large losses in total lipolytic activity had occurred, as after treatment with a mixture of ether, ethanol and deoxycholate, or prolonged action of trypsin. When the concentration of taurodeoxycholate was increased from zero to 1 mM in a tributyrin substrate, the lipolytic activites of calf salivary and pancreatic lipases, and pig pancreatic lipase, increased. At a concentration of 4 mM-taurodeoxycholate, calf salivary lipase activity was higher, that of calf pancreatic lipase slightly lower and pig pancreatic lipase activity markedly lower.

USE OF GEL FILTRATION AND ION EXCHANGE CHROMATOGRAPHY FOR PARTIAL FRACTIONATION OF A SOLUTION FROM WHEY CONTAINING SODIUM CHLORIDE, ASH AND LACTOSE

1971 ◽  
Vol 34 (5) ◽  
pp. 241-243
Author(s):  
M. A. Khorshid ◽  
I. D. Rifaat ◽  
M. H. Abd El-Salam ◽  
A. A. Hofi
Keyword(s):  
Sodium Chloride ◽  
Ion Exchange ◽  
Gel Filtration ◽  
Chloride Content ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
The Other ◽  
Double Pass ◽  
Pass System ◽  
Other Hand

Lactose and chloride are partially separated from deproteinated salted whey (D.S.W.) by gel filtration. Approximately 24.1% of chloride was removed and 8.9% lactose was recovered from 3 ml of D.S.W. with a 10 g column of Sephadex G-10. On the other hand, 77.6% of chloride was removed and 74.5% lactose was recovered from 3 ml of D.S.W. with three continuous 10 g columns of Sephadax G-10. Ion exchange chromatography removed 92.1% and 99.1% of the chloride content of D.S.W. by using a single and double pass system, respectively.

Caractéristiques fonctionnelles des activités peroxydasiques des feuilles et cals d'une plante à métabolisme acide crassulacéen, Pedilanthus tithymaloides variegatus, Euphorbiaceae

1984 ◽  
Vol 62 (9) ◽  
pp. 901-907 ◽  
Author(s):  
Pierre Bricage
Keyword(s):  
Ion Exchange ◽  
Physicochemical Properties ◽  
Affinity Chromatography ◽  
Enzyme Activities ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Leaf Tissues ◽  
Specific Activities ◽  
Isoenzyme Patterns

Peroxidases (donor:hydrogen-peroxide oxidoreductase; EC 1.11.1.7) from leaf tissues and calli were extracted and compared in terms of their specific activities at different pH and temperatures, their isoenzyme patterns and physicochemical properties. Three groups of enzyme activities were detected and their purification was performed by conventional methods (ammonium sulfate fractionation, ion-exchange chromatography, gel filtration, affinity chromatography).

Studies on an Inhibitor of Plasminogen Activation in Human Serum

1973 ◽  
Vol 30 (02) ◽  
pp. 414-424 ◽  
Author(s):  
Ulla Hedner
Keyword(s):  
Ion Exchange ◽  
Human Serum ◽  
Antithrombin Iii ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Specific Adsorption ◽  
Partial Purification ◽  
Plasminogen Activation ◽  
Preparative Electrophoresis

SummaryA procedure is described for partial purification of an inhibitor of the activation of plasminogen by urokinase and streptokinase. The method involves specific adsorption of contammants, ion-exchange chromatography on DEAE-Sephadex, gel filtration on Sephadex G-200 and preparative electrophoresis. The inhibitor fraction contained no antiplasmin, no plasminogen, no α1-antitrypsin, no antithrombin-III and was shown not to be α2 M or inter-α-inhibitor. It contained traces of prothrombin and cerulo-plasmin. An antiserum against the inhibitor fraction capable of neutralising the inhibitor in serum was raised in rabbits.

scholarly journals Isolation, partial purification and characterization of phospholipase A2 from Naja Katiensis venom

2021 ◽  
Vol 13 (2) ◽  
pp. 107-112
Author(s):  
C.F. Okechukwu ◽  
P.L. Shamsudeen ◽  
R.K. Bala ◽  
B.G. Kurfi ◽  
A.M. Abdulazeez
Keyword(s):  
Ion Exchange ◽  
Phospholipase A2 ◽  
Specific Activity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Partial Purification ◽  
Crude Venom

The most effective and acceptable therapy for snakebite victims is the immediate administration of antivenin which is limited by problems of hypersensitivity reactions in some individuals and its inability to resolve the local effects of the venom. The aim of this study was to isolate, partially purify and characterize phospholipase A2 from Naja Katiensis venom. Phospholipase A2 was partially purified via a two-step process: gel filtration on Sephadex G-75 and ion exchange chromatography using CM Sephadex, and subjected to SDS-PAGE analysis. From the results, the specific activity of the partially purified PLA2 decreased from 0.67μmol/min/mg in crude venom to 0.29μmol/min/mg after ion exchange chromatography with a yield of 5% and purification fold of 0.43. The optimum temperature of the purified PLA2 was found to be 35ºC and optimum p.H of 7. velocity studies for the determination of kinetic constants using L-a-lecithin as substrate revealed a Km  of 1.47mg/ml and Vmax  of 3.32μ moles/min/mg. The sodium dodecyl sulphate polyacrylamide gel electrophoresis of the purified PLA2 showed a distinct band with molecular weight estimated to be 14KDa. In conclusion, the present study shows that phospholipase A2 was isolated, purified and characterized. This may serve as a promising candidate for future development of a novel anti-venin drug.

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