Desorption, biospecific

Thus a biospecific adsorbant was obtained selective desorption should therefore permit successive elution of cellulolytic enzymes. It was thus found that the method was very useful in the purification of the cellobio-hydrolases from Tr. r., starting from very crude culture filtrates. Addition of glucose (0.1M) or gluconolactone suppressed the action of glucosidases present, preventing deterioration of the columns. The capacities exceeded 10 mg CBH I per ml gel (prepared with CNBr activated Sepharose). 

Ion exchange chromatography, hydrophobic interaction chromatography (HlC), and affinity chromatography (AC) show some similarities concerning practical realization therefore, most of the hints given for lEC are applicable for HlC and AC. Examples for AC are given in Protocols 3.6.2.4 (biospecific desorption) and 3.6.2.5 (elution by partial denaturation). 

Competitive desorption (elution) has the advantage of very smooth conditions for protein structure, but the disadvantage of difficult removing of the ligand when further biospecific interactions are intended. 

This allowed a biospecific desorption procedure using AT III, heparin (Hep) and Hep-AT III complex. The present work provides further... 

Desorption of Bound Thrombin by Antithrombin III, Heparin or Hep-AT III Complex. Thrombin was injected onto PAOM or PSSO chromatographic supports in 0.1 M NaCl solution. To minimize possible kinetic effects, the flow-rate was reduced to 0.2 ml/min. After washing with 0.1 M NaCl buffer, an excess of either AT III, Hep, or AT Ill-Hep complex was injected. The flow was then stopped for five minutes in order to obtain a better exchange of macromolecules between the mobile and the stationary phases. The objective was to determine whether AT III, Hep or AT Ill-Hep complex could act as eluents for the "biospecific desorption" of the bound enzyme at 0.1 M NaCl. 

For the separation biospecific affinity chromatography was used, where in addition to the adsorption and desorption zone a washing zone has to be included. Gottschlich and Kasche [22] used the same apparatus for the separation of monoclonal antibodies from a fermentation broth. 

Affinity chromatography Biospecific adsorption/desorption Molecular structure... 

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