Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.2.1.23 (beta-galactosidase)
 14,648 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

This paper describes a theoretical approach to the prediction of the performance of preparative affinity separations for biological macromolecules in packed columns. The approach, which is applicable to conventional low-pressure packed column methods as well as high-performance liquid chromatography techniques, requires knowledge of certain parameters that describe the interactions between adsorbent and adsorbate during the affinity separation procedure. We have measured the parameters appropriate to the adsorption stages of affinity systems involving immobilised Cibacron Blue and immobilised monoclonal antibodies against beta-galactosidase. The theoretical predictions appear to agree well with the experimental performance of batch and packed column affinity systems. The influence of the factors that govern the performance of the adsorption stage of the separation procedure is explained in detail, and the possible advantages of using HPLC techniques in macropreparative affinity chromatography are discussed.
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PMID:Prediction of the performance of preparative affinity chromatography. 643 81

Continuous superporous agarose beds constitute a new support material for chromatography, biocatalysis and electrophoresis. The bed consists of a single piece of agarose gel, homogeneously transected by flow-carrying pores, which easily can be varied in the range of 10-100 microm. In this work, large diameter beds (60 mm) were prepared and used in specially designed radial flow columns. The basic chromatographic properties of the beds were investigated by size-exclusion chromatography experiments. In an affinity chromatography application one bed was derivatized with Cibacron Blue 3GA and used for the purification of lactate dehydrogenase from a crude bovine heart extract. In a biotransformation application one bed was provided with immobilized beta-galactosidase and used in the production of lactose-free milk.
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PMID:Continuous superporous agarose beds in radial flow columns. 1151 18

To obtain silica supports for high-performance affinity chromatography, a method of preparing CNBr-activated diol-silica under anhydrous conditions was developed. Activation of the silane-derived hydroxyls with cyanogen bromide and triethylamine was optimized and demonstrated to efficiently couple several amino ligands (tryptophan, 6-aminohexyl-Cibacron Blue, and DNA). Sonication and vacuum degassing, a procedure used to remove air from the silica beads, increased activation. Coupling of an amino ligand under slightly basic conditions (pH 8.0) gave the highest yield. The linkage between the immobilized ligands and silica was stable from pH 2-10. Anhydrous acetone was the most effective solvent for activation but dimethylformamide and 2-propanol were also good choices. The high-performance affinity chromatography columns obtained by coupling sequence-specific DNA binding sequences for Lac repressor-beta-galactosidase fusion protein were compared to affinity columns obtained by coupling the same DNA element to Sepharose beads; 5 microm silica gave the best performance.
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PMID:Cyanogen bromide activation and coupling of ligands to diol-containing silica for high-performance affinity chromatography optimization of conditions. 1235 Jan 29