Agarose immobilization

Kriz D, Mosbach K. Competitive amperometric morphine sensor-hased on an agarose immobilized molecularly imprinted polymer. Anal Chim Acta 1995 300 71-75. 

Unless otherwise stated, enzymes were immobilized on agarose. Immobilized in PAN gel Immobilized on silica gel-glutaialdehyde (a six-fold excess of CTP was utilized). 

Another commonly used reaction for insolubilization of proteins and ligands through amino groups to polysaccharides is with 2,4,6-trichlorotriazine [18,19]. This procedure is extensively used in our laboratories in the preparation of agarose immobilized enzymes. 

Rp protein immobilization yield (mg immobilizEd enzyme/mg contacted protein) Re enzyme immobilization yield (units of enzyme activity expressed in the biocatalyst/unit of enzyme activity contacted) SA specific activity of the biocatalyst (units of enzyme activity expressed/unit mass of biocatalyst). OIL octyl agarose immobilized lipase BIL butyl Sepabeads immobilized lipase... 

Investigation of the binding characteristics of urinary acid 3-D-gaIactosidase, 3-D-2-acetamido-2-deoxyglucosidase, a-D-galactosidase and a-L-fucosidase from patients with mucolipidosis II and mucolipidosis III to agarose-immobilized concanavalin A revealed a two to ten-fold decrease in the proportion of enzyme (that adsorbed on the lectin) activities from patients with mucolipidoses II and III. Neuraminidase treatment of the unabsorbed enzyme fraction did not... 

Kriz, D. Mosbach, K. Competitive Amperometric Morphine Sensor Based on an Agarose Immobilized Molecularly Imprinted Polymer. Anal. Chim. Acta 1995, 300, 71-75. 

Antiserum - protein A agarose (immobilized antibody for boronate chromatography)... 

In normal and regenerating rat liver tissue, /8-D-2-acetamido-2-deoxygluco-sidase has been shown to be localized primarily in the fraction of light mitochondria, enriched with lysosomes. The interaction of rat liver )8-d-2-acetamido-2-deoxyhexosidase with agarose-immobilized ovalbumin has been investigated. The use of the latter as an aflfinity chromatography material was also studied. 

Human serum a-L-fucosidase has been purified 240 000 fold with 35 % yield by an affinity chromatographic procedure utilizing agarose-immobilized 2-amino-2-deoxy-L-fucose. Isoelectric focusing of the purified enzyme indicated the presence of several forms (major form p/ 5.0). Sodium dodecyl sulphate-polyacrylamide gel electrophoresis of the enzyme (mol. wt. 2.96 0.30 X 10 by gel filtration) indicated that the presence of two subunits with similar molecular weights (5.65 x 10 and 5.40 x 10 ). Using the 4-nitro-phenyl substrate (Xm value 0.52 mM) the enzyme displayed a broad pH optimum centred round pH 4.8 with a second minor optimum at pH 6.1. The enzyme was shown to contain 1.7% neuraminic acid quantitative immuno-precipitation studies were also conducted on the enzyme. 

A simple procedure reported for the isolation of jack-bean a-D-mannosidase was based on affinity chromatography on agarose-immobilized benzidine. The influence of substituents on the hydrolysis of substituted phenyl a-D-mannopyranosides by a-D-mannosidase from Medicago sativa seeds has been investigated. As indicated by structure-activity relations, the electronic effect of the substituent has an influence on the rate of formation of the intermediate D-mannosyl-enzyme complex. This effect depends not only on the nature of the substituent, but also on its position meta or para) and on the temperature of the experiment. Hammett-type linear free energy relationships show that the reaction constant p changes its sign at ca. 27 °C. Substrates... 

A simple procedure described for the detection of encarbohydrate moiety of ribonuclease B has a-D-mannopyranosyl residues as terminal non-reducing groups, but such residues are lost as part of an oligosaccharide on action of the endo -o-l-acet-amido-2-deoxyglucanase, agarose immobilized concanavalin A was used to separate radioactive substrate (absorbed) from radioactive product (unadsorbed). The assay was used to follow the preparation of the enzyme from Streptomyces plicatus. 

The purification of pullulanase from Aerobacter aerogenes via affinity chromatography on an agarose-immobilized cyanogen bromide-activated cyclohexa-amylose has been reported. Elution from the affinity column was achieved using cyclohepta-amylose. Crossed immunoelectrophoresis was used to follow the various steps of the purification. 

The arylsulphatases A and B of human urine, rat liver, rat brain, and sheep brain have been separated by a simple and rapid affinity chromatographic method on agarose-immobilized Cibacron Blue F3GA. ... 

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