This paper describes a simple purification procedure for protease nexin, a serine proteinase inhibitor secreted by cultured human fibroblasts that regulates proteinase activity at and near the cell surface. The first step in the procedure takes advantage of the high-affinity binding of protease nexin to dextran sulphate-Sepharose. This step eliminates the need for prior concentration of the ser...

Bandeiraea simplicifolia I (BS I) isolectins, immobilized on Sepharose beads, specifically retarded low molecular weight ligands containing terminal cu-o-galactopyranosyl and 2acetamido-2-deoxy-a-D-galactopyranosyl residues. A BS I lectin-Sepharose column has been used to perform very efficient separations of structurally homologous sugar nucleotides and oligosaccharides. For example, UDP-gluco...

The entomopathogenic fungus Verticillium lecanii is a well-known biocontrol agent of fungal phytopathogens, as well as insect pests. A 42-kDa chitinase belonging to family 18 of the glycosyl hydrolases was isolated and partially characterized. Chitinase was purified using successive column chromatography on phenyl-sepharose, DEAE-sepharose, and CM-sepharose. The enzyme showed the highest activi...

The interactions of Sepharose 4B-immobilized concanavalin A (ConA) with 10 glycoasparagines derived from ovalbumin were investigated quantitatively by frontal affinity chromatography. In this method, a carbohydrate solution is applied continuously to a ConA-Sepharose column and the retardation of the elution front is measured as a parameter of the strength of the interaction. The dissociation c...

Extraction, concentration, and serological detection of staphylococcal enterotoxins from foods are laborious and time consuming. By exposing food extracts to an insoluble matrix tagged with specific anti-enterotoxin B, we have been able to recover the toxin from foods in a sensitive and rapid way. After mixing the reagents for 2 h at room temperature, immunoglobulin G antibodies were attached t...

Cultured human monocytes released cytostatic activity upon in vitro activation with lymphokines and lipopolysaccharide. This activity was mainly due to the presence of two different cytostatic factors, termed CstF I and II, which were separated by ion-exchange chromatography. At neutral pH, CstF I bound to the weak anion exchanger DEAE-Sephacel but not to the weak cation exchanger CM-Sepharose,...

We describe here a simple, rapid chromatographic procedure for quantitatively separating serum creatine kinase isoenzymes (EC 2.7.3.2), with diethylaminoethyl (DEAE)-Sepharose CL-6B as the anion-exchanger. We established the column bed height and the elution parameters by use of a simplex procedure. DEAE-Sepharose CL-6B equilibrated in tris(hydroxymethyl)aminomethane (50 mmol/L, pH 7.5, and con...

A commercial and very hydrophobic styrene-divinylbenzene matrix, MCI GEL® CHP20P, has been compared to octyl-Sepharose® beads as support to immobilize three different enzymes: lipases from Thermomyces lanuginosus (TLL) and from Rhizomucor miehie (RML) and Lecitase® Ultra, a commercial artificial phospholipase. The immobilization mechanism on both supports was similar: interfacial activation of ...

The capacities of Procion Red HE-3B and Cibacron Blue F3G-A immobilized to Sepharose CL-4B and Matrex 201R for NAD+-, NADP+- and NAD(P)+-dependent dehydrogenases were measured. Procion Red HE-3B columns retarded NADP+-dependent dehydrogenases more effectively than NAD+-dependent dehydrogenases, whilst immobilized Cibacron Blue F3G-A retarded NAD+-dependent dehydrogenases more effectively than N...