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.

Purification of glysojanin, an antifungal protein, from the black soybean Glycine soja

2003 ◽  
Vol 81 (6) ◽  
pp. 387-394 ◽  
Author(s):  
Patrick H.K Ngai ◽  
T B Ng
Keyword(s):  
Liquid Chromatography ◽  
Ion Exchange ◽  
Glycine Soja ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Fast Protein Liquid Chromatography ◽  
Antifungal Protein ◽  
Black Soybean

A monomeric protein, with a molecular mass of 25 kDa and an N-terminal sequence resembling a segment of chitin synthase, was isolated from the seeds of the black soybean Glycine soja. The protein, designated glysojanin, demonstrated potent antifungal activity against the fungi Fusarium oxysporum and Mycosphaerella arachidicola. It inhibited HIV-1 reverse transcriptase with an IC50 of 47 µmol/L, [methyl-3H]thymidine incorporation by mouse spleen cells with an IC50 of 175 µmol/L, and translation in the rabbit reticulocyte lysate with an IC50 of 20 µmol/L. Glysojanin was purified using a procedure that involved ion-exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel blue gel, ion-exchange chromatography by fast protein liquid chromatography on Mono S, and gel filtration by fast protein liquid chromatography on Superdex 75.Key words: antifungal protein, seeds, soybean, purification.

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.

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).

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.

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.

Preparation and properties of three analogues of [5-leucine]enkephalin, substrates for enzyme conversion studies

1985 ◽  
Vol 50 (6) ◽  
pp. 1329-1334
Author(s):  
Jaroslav Vičar ◽  
Linda Servítová ◽  
Martin Flegel ◽  
Karel Hauzer ◽  
Tomislav Barth
Keyword(s):  
High Pressure ◽  
Liquid Chromatography ◽  
Guinea Pig ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Guinea Pig Ileum ◽  
Opioid Activity ◽  
C Terminus ◽  
N Terminus ◽  
Fragment Condensation

Analogues of [5-Leu]enkephalin, prolonged by methionine on the N-terminus or, by lysine or methionine on the C-terminus were prepared by fragment condensation, purified by ion exchange chromatography or high-pressure liquid chromatography. The substances were characterised by their opioid activity in a test on guinea-pig ileum in comparison with the activity of [5-Leu]enkephalin.

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.

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