Isolation of factor IX concentrates for clinical use by ion-exchange chromatography and ammonium sulphate precipitation

1986 ◽  
Vol 363 (1) ◽  
pp. 95-100 ◽  
Author(s):  
D. Stampe ◽  
B. Wieland ◽  
A. Köhle
Keyword(s):  
Ion Exchange ◽  
Ammonium Sulphate ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Factor Ix ◽  
Clinical Use ◽  
Ammonium Sulphate Precipitation

PURIFICATION AND CHARACTERISATION OF FIBRINOLYTIC ENZYMES FROM ENDOPHYTIC FUNGI AND LIGNOSUS RHINOCERUS

2016 ◽  
Vol 78 (6-5) ◽  
Author(s):  
Zainon Mohd Noor ◽  
Mohd Sidek Ahmad ◽  
Zaidah Zainal Ariffin
Keyword(s):  
Ion Exchange ◽  
Endophytic Fungi ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Fibrinolytic Enzymes ◽  
Molecular Weights ◽  
Ammonium Sulphate Precipitation ◽  
Fusarium Sp ◽  
Different Sources

Three enzymes FH3, S13 and LR1 from three different sources showed fibrinolytic activities. Two were from endophytic fungal cultures and one from the sclerotium of Lignosus rhinocerus mushroom (LR1). FH3, S13 cultures and LR1, the crude extract of the sclerotium were concentrated and purified by ammonium sulphate precipitation, ion-exchange chromatography and gel-filtration. The molecular weights of the FH3, S13 and LR1 purified enzymes were estimated to be approximately 34kDa, 34kDa and 10kDa, respectively. Maximum fibrinolytic activities were observed for FH3 at pH 7 and 30°C, S13 at pH 8 and 40°C and LR1 at pH 6 and 40°C.  In our earlier paper we identified FH3 as Fusarium sp. and S13 as Penicilium citrinum. 

LINGCOD MUSCLE PHOSPHORIBOISOMERASE AND RIBULOSE 5′-PHOSPHATE 3′-EPIMERASE

1959 ◽  
Vol 37 (8) ◽  
pp. 961-973 ◽  
Author(s):  
H. L. A. Tarr
Keyword(s):  
Acid Phosphatase ◽  
Ion Exchange ◽  
Ammonium Sulphate ◽  
Simple Procedure ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Water Extraction ◽  
Enzyme Preparations

Comparatively pure phosphoriboisomerase and ribulose 5′-phosphate 3′-epimerase enzyme preparations were obtained from lingcod muscle by a simple procedure involving water extraction, saturation of the extract with ammonium sulphate, dialysis, brief heating to 55 °C, lyophilization of the solution, and final separation by ion exchange chromatography, using diethylamiuoethyl cellulose columns. Both enzymes have broad pH optima above pH 7.0, but are rapidly inactivated below this value. The following equilibria were established and compared with those obtained by other investigators: [Formula: see text], 1.35:1.0; [Formula: see text], 1: 1.5 and [Formula: see text], 1:0.58:0.66. The ketopentulose phosphate resulting from the action of phosphoriboisomerase on D-ribose 5-phosphate was isolated and identified as D-ribulose5-phosphate. Both D-ribulose and D-xylulose were demonstrated after subjecting a product of epimerase action to hydrolysis by acid phosphatase and ion exchange chromatography.

LINGCOD MUSCLE PHOSPHORIBOISOMERASE AND RIBULOSE 5′-PHOSPHATE 3′-EPIMERASE

1959 ◽  
Vol 37 (1) ◽  
pp. 961-973
Author(s):  
H. L. A. Tarr
Keyword(s):  
Acid Phosphatase ◽  
Ion Exchange ◽  
Ammonium Sulphate ◽  
Simple Procedure ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Water Extraction ◽  
Enzyme Preparations

Comparatively pure phosphoriboisomerase and ribulose 5′-phosphate 3′-epimerase enzyme preparations were obtained from lingcod muscle by a simple procedure involving water extraction, saturation of the extract with ammonium sulphate, dialysis, brief heating to 55 °C, lyophilization of the solution, and final separation by ion exchange chromatography, using diethylamiuoethyl cellulose columns. Both enzymes have broad pH optima above pH 7.0, but are rapidly inactivated below this value. The following equilibria were established and compared with those obtained by other investigators: [Formula: see text], 1.35:1.0; [Formula: see text], 1: 1.5 and [Formula: see text], 1:0.58:0.66. The ketopentulose phosphate resulting from the action of phosphoriboisomerase on D-ribose 5-phosphate was isolated and identified as D-ribulose5-phosphate. Both D-ribulose and D-xylulose were demonstrated after subjecting a product of epimerase action to hydrolysis by acid phosphatase and ion exchange chromatography.

scholarly journals Purification and Characterization of Cellulase from Termite Ametermes eveuncifer (Silverstri) Soldiers

2016 ◽  
Vol 9 (1) ◽  
pp. 1 ◽  
Author(s):  
B. S. Fagbohunka ◽  
R. E. Okonji ◽  
Ayinla Zainab Adenike
Keyword(s):  
Ion Exchange ◽  
Specific Activity ◽  
Ion Exchange Chromatography ◽  
Maximum Activity ◽  
Exchange Chromatography ◽  
Purification And Characterization ◽  
Ammonium Sulphate Precipitation ◽  
Percentage Yield ◽  
Termite Soldiers

Cellulase enzyme was purified and characterized from termite soldiers (Ametermes eveuncifer) using 70% ammonium sulphate precipitation, ion exchange chromatography and affinity chromatography. The enzyme isolated had a specific activity of 5.04 U/mg with a percentage yield of 11.7%. The enzyme showed maximum activity at 500C and pH 8. The enzyme was not inhibited by Ba2+ at a concentration of 1mM and Pb2+ at 10 mM concentration but was inhibited by other metal ions at 1 mM and 10 mM concentrations of their salts (NaCl, KCl, MnCl2, and NiCl2),

Rapid purification of factor IX, factor X and prothrombin by immunoaffinity and ion exchange chromatography

1989 ◽  
Vol 55 (1) ◽  
pp. 121-133 ◽  
Author(s):  
Syed S. Ahmad ◽  
Razia Rawala-Sheikh ◽  
Arthur R. Thompson ◽  
Peter N. Walsh
Keyword(s):  
Ion Exchange ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Factor Ix ◽  
Factor X ◽  
Rapid Purification

scholarly journals PEMURNIAN ENZIM SEFALOSPORIN-C ASILASE DAN OPTIMASI PROSES KROMATOGRAFI PENUKAR ION

2018 ◽  
Vol 5 (2) ◽  
pp. 119
Author(s):  
Uli Julia Nasution ◽  
Silvia Melinda Wijaya ◽  
Ahmad Wibisana ◽  
Anna Safarrida ◽  
Indra Rachmawati ◽  
...  
Keyword(s):  
Ion Exchange ◽  
Ammonium Sulphate ◽  
Specific Activity ◽  
Exchange Chromatography ◽  
Size Exclusion ◽  
Cephalosporin C ◽  
Α Subunit ◽  
Ammonium Sulphate Precipitation ◽  
Cephalosporin C Acylase ◽  
Sds Page

Purification of Cephalosporin-C Acylase and Its Optimization of Ion-Exchange ChromatographyABSTRACTCephalosporin-C acylase (CCA) has an important role in the one-step conversion of cephalosporin-C into 7-ACA. Purification process aims to increase specific activity of CCA enzyme. Purification began with cell lysis, ammonium sulphate precipitation, dialysis, ion exchange chromatography (IEC) and size exclusion chromatography. IEC optimization of elution step was also done to compare gradient and isocratic elusion. Purification was capable to increase the enzyme purity upto 33.66 fold, with specific activity of 3.00 U/mg and the yield reached 41.41%. Optimization of elusion during IEC showed that isocratic protein elusion was more efficient (taking shorter time, 3 column volume (CV) only) than that of gradient batch (up to 9 CV). SDS-PAGE analysis demonstrated that the recombinant CCA enzyme existed in two types, active enzyme containing α-subunit (25 kDa) and β-subunit (58 kDa), and inactive enzyme (83 kDa) as precursor. Furthermore, 30% ammonium sulphate saturated precipitation was able to precipitate this inactive CCA.Keywords: 7-ACA,CCA, cephalosporin C, protein purification, specific activity ABSTRAKSefalosporin-C asilase (CCA) merupakan enzim yang berperan penting dalam konversi satu tahap sefalosporin-C menjadi 7-ACA. Proses purifikasi merupakan salah satu cara untuk meningkatkan aktivitas spesifik enzim CCA. Proses purifikasi dimulai dari memecah sel, diikuti dengan tahap presipitasi menggunakan amonium sulfat, dialisis, kromatografi penukar ion (IEC) dan kromatografi eksklusi. Optimasi proses IEC pada tahap elusi juga dilakukan untuk membandingkan elusi enzim CCA secara gradien dan isokratik. Proses purifikasi pada penelitian ini mampu meningkatkan kemurnian enzim hingga 33,66 kali, dengan aktivitas spesifik sebesar 3,00 U/mg dan perolehan enzim sebesar 41,41%. Hasil optimasi IEC pada proses elusi secara isokratik lebih efisien dari segi waktu (hanya membutuhkan 3 kolom volume (CV) dibandingkan dengan secara gradien (sampai 9 CV). Hasil SDS-PAGE menunjukkan bahwa CCA rekombinan merupakan enzim dengan 2 macam bentuk yaitu enzim aktif, yang terdiri dari subunit α (25 kDa) dan β (58 kDa), dan enzim tidak aktif berupa prekursor (83 kDa). Proses presipitasi menggunakan amonium sulfat 30% tersaturasi dapat mengendapkan prekursor CCA.Kata Kunci: 7-ACA, aktivitas spesifik, CCA, purifikasi protein, sefalosporin C

scholarly journals Biochemical and Kinetic Study for the partially purified Lecithin: Cholesterol acyltransferase from serum cardiovascular disease

2020 ◽  
Vol 25 (2) ◽  
pp. 31
Author(s):  
Amel Taha Yassein Al-Juraisy1 ◽  
Ameera Aziz Mahmood Al-Juraisy1 ◽  
Ameera Aziz Mahmood Al-Juraisy1 ◽  
Nadia Ahmed Saleh2
Keyword(s):  
Cardiovascular Disease ◽  
Molecular Weight ◽  
Ion Exchange ◽  
Cholesterol Acyltransferase ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Partial Purification ◽  
Lecithin Cholesterol Acyltransferase ◽  
Ammonium Sulphate Precipitation ◽  
Optimum Ph

The study includes partial purification of Lecithin: Cholesterol acyltransferase (LCAT) from the blood serum of a person suffering from atherosclerosis. Several techniques were applied including ammonium sulphate precipitation, dialysis, ion exchange chromatography and electrophoresis. It was found that LCAT has one isoenzyme and the highest activity was (1073.46 × 10-3) unit/ml and a molecular weight of 62 KDa. The study  also deals with characterizing of LCAT. It was found that the optimum pH and temp. of the enzyme were 7 and 35ºC.    http://dx.doi.org/10.25130/tjps.25.2020.027

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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