Interference with carcinoembryonic antigen radioimmunoassays by glycosaminoglycans, and their removal.

1983 ◽  
Vol 29 (12) ◽  
pp. 2049-2053 ◽  
Author(s):  
J T Wu ◽  
E Mau ◽  
J A Knight
Keyword(s):  
Ion Exchange ◽  
Carcinoembryonic Antigen ◽  
Plasma Proteins ◽  
Acid Treatment ◽  
Uronic Acid ◽  
Anticoagulant Activity ◽  
Gel Filtration ◽  
The Other ◽  
Negative Effects ◽  
Ion Exchange Column

Abstract Using the reagents from the CEA-Roche kit, we found that solutions containing only glycosaminoglycans (GAG) yielded false concentrations of carcinoembryonic antigen (CEA), and mixtures of CEA and GAG produced falsely high values. On the other hand, solutions of GAG yielded no additional CEA concentration when reagents from the Abbott CEA kit were used; rather, the CEA result was decreased with the Abbott kit for a CEA solution also containing GAG. These effects of GAG were not ascribable to contamination, because neither gel filtration nor ion-exchange column chromatography separated the false Roche CEA content related to GAG from their peaks of uronic acid or from their anticoagulant activity. In addition, an enzyme specific for GAG eliminated these GAG-derived false concentrations. Both the positive and negative effects are correlated with concentrations of GAG. We found that the concentration of GAG could be decreased in a solution containing plasma proteins by either heating (70 degrees C, 15 min) or treating with perchloric acid (0.6 mol/L). The former is superior because essentially all GAG added to a solution containing plasma proteins were removed by heat, whereas as much as 25% to 80% of the GAG still remained after acid treatment. The effect of GAG was also completely eliminated by treating the specimens with chondroitinase ABC lyase (EC 4.2.2.4).

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.

Purification and Identification of Lactoferrin from Bovine Milk

2012 ◽  
Vol 524-527 ◽  
pp. 2290-2293
Author(s):  
Ying Ying Kong ◽  
Meng Liu ◽  
Wei Di ◽  
Cong Wang ◽  
Ming Du ◽  
...  
Keyword(s):  
Ion Exchange ◽  
Membrane Filtration ◽  
Bovine Milk ◽  
Gel Filtration ◽  
Exchange Column ◽  
Sds Page ◽  
Ion Exchange Column

Lactoferrin has many kinds of bioactivities which have attracted more and more attention. In the present study, lactoferrin from bovine milk was isolated and purified by membrane filtration, series of chromatography on SP Sepharose Big Bead ion exchange column and Superdex 200 gel filtration column. The purified lactoferrin was identified by SDS-PAGE compared with the lactoferrin standard.

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 .

scholarly journals The xylanase system of Coniophora cerebella

1968 ◽  
Vol 108 (4) ◽  
pp. 571-576 ◽  
Author(s):  
N. J. King ◽  
D. B. Fuller
Keyword(s):  
Molecular Weight ◽  
Culture Filtrate ◽  
Uronic Acid ◽  
Enzyme System ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
The Other ◽  
Xylose Residue ◽  
Random Manner ◽  
Acidic Sugars

1. The culture filtrate of the fungus Coniophora cerebella grown on poplar 4-O-methylglucuronoxylan as carbon source and enzyme inducer contained an enzyme system that degraded the polysaccharide to xylose, acidic and neutral oligosaccharides and an enzyme-resistant polymer. Free uronic acid was not produced. 2. Cold ethanol fractionation of the culture filtrate yielded two active fractions, one of which had only xylanase (EC 3.2.1.8) and the other both xylanase and xylosidase (EC 3.2.1.37) activities. Further fractionation on DEAE-cellulose resolved the xylanase and xylosidase activities. 3. The xylanase degraded poplar 4-O-methylglucuronoxylan in an essentially random manner, producing oligosaccharides, but some xylose residues in the vicinity of uronic acid side groups were protected from hydrolysis, preventing a truly random attack. The xylosidase attacked the polysaccharide very slowly, releasing xylose, but the oligosaccharides produced by the action of the xylanase were much more susceptible to hydrolysis by the xylosidase. 4. The products of xylanase action were separated into neutral and acidic fractions. The neutral oligosaccharides were separated by chromatography on charcoal–Celite, and the major products were characterized as xylobiose, xylotriose, xylotetraose and xylopentaose. Some of the acidic sugars were branched, having the uronic acid residue attached to a xylose residue other than the terminal non-reducing one. 5. Gel filtration of various xylanase fractions gave values for the molecular weight of the enzyme from 34000 to 38000.

scholarly journals PURIFICATION AND CHARACTERIZATION OF POLYGALACTURONASE OF `FUJI' APPLES.

HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 653e-653
Author(s):  
Seung-Ryeul Shin ◽  
Jae-Kyun Byun ◽  
Kyung-Ho Chang
Keyword(s):  
Ion Exchange ◽  
Column Chromatography ◽  
Malus Domestica ◽  
Gel Filtration ◽  
Reducing Sugar ◽  
Purification And Characterization ◽  
Stable Temperature ◽  
Ion Exchange Column ◽  
Optimum Ph

Polygalacturonase (PG) was purified from apple, Malus domestica Borkh, cv. Fuji by gel filtration, CM cellulose ion exchange column chromatography and characterized by means of several biochemical methods. Two forms of isozymes, PG-I and PG-II, were detected and the activities of PG-I were found to be higher than PG-II. The Km and Vmax values were calculated to be 1.54 mg/ml and 0.25 μM with reducing sugar 1 ml/30min., respectively. The PG was active between pH 3 and 8 with the optimum pH of about 4-5. The stable temperature for the PG was below 55°C with 30°C optimum. The PG activities were increased by Na* and Cu**, but were inhibited by Ag*, EDTA and SDS.

In Vitro Anticoagulant Activity of Esters of Heparin.

1979 ◽  
Author(s):  
J. Mardiguian ◽  
M. Trillou ◽  
J. Marin
Keyword(s):  
Molecular Weight ◽  
Ion Exchange ◽  
Chemical Modification ◽  
Anticoagulant Activity ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Hydroxyl Groups ◽  
Oh Groups

Chemical modification of Heparin by esterification of its hydroxyl groups has been carried out in order to study its influence on the in vitro anticoagulant activity. Beef and pig mucosal Heparins have been fractionated by gel filtration and by ion-exchange chromatography to yield fractions which have been esterified.Two types of esters have been prepared : 4-chlorophenoxy-acetic and 4-chlorophenoxy-isobutyric.The anticoagulant activity of the these esters has been evaluated in vitro (using U.S.P. method, anti-Xa and anti-thrombin assays) and correlated with their chlorine content and U.V. absorption.This study shows that the in vitro anticoagulant activity of heparin is decreased by esterification of its OH groups, to an extent depending upon the type of ester, the molecular weight and the degree of substitution.The results of these experiments will be reported and discussed in detail.

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.

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.

Purification of the Antihemophilic Factor by Gel Filtration on Agarose

1969 ◽  
Vol 22 (03) ◽  
pp. 577-583 ◽  
Author(s):  
M.M.P Paulssen ◽  
A.C.M.G.B Wouterlood ◽  
H.L.M.A Scheffers
Keyword(s):  
Factor Viii ◽  
Plasma Proteins ◽  
Large Scale ◽  
Gel Filtration ◽  
Large Scale Preparation ◽  
Scale Preparation ◽  
Antihemophilic Factor

SummaryFactor VIII can be isolated from plasma proteins, including fibrinogen by chromatography on agarose. The best results were obtained with Sepharose 6B. Large scale preparation is also possible when cryoprecipitate is separated by chromatography. In most fractions containing factor VIII a turbidity is observed which may be due to the presence of chylomicrons.The purified factor VIII was active in vivo as well as in vitro.

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.

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