CNBr-activated Sepharose™ Fast Flow resin is an affinity chromatography resin designed for multipoint attachment of protein ligands containing large amino groups. The laboratory-scale use of CNBr-activated coupling media based on Sepharose™ resin is very well-documented. However, detailed information about process-scale use and manufacturing applications of CNBr-activated Sepharose™ 4 Fast Flow resin is generally not available. Nevertheless, the use of this medium in the development of a process to purify native gp120 from HIV-1 infected T-cells has been described (see data file for reference and details). Here, the authors coupled Galanthus nivalis agglutinin (GNA), a lectin from the bulb of the snowdrop, to CNBr-activated Sepharose™ 4 Fast Flow resin and then used the coupled gel to help purify the outer envelope glycoprotein gp120 of HIV-1, which is a major target for immunotherapy.

The coupling reaction, which is rapid and spontaneous, is easy to carry out and requires no special chemicals or equipment. CNBr-activated Sepharose™ 4 Fast Flow resin is supplied as a freeze-dried powder stabilized with additives. Coupling a ligand to the activated matrix involves first swelling and washing the gel followed by coupling. Instructions included with the product describe methods for coupling ligands and the effect of different conditions on the coupling efficiency. Users should develop a specific procedure for each individual application. See data file for more information.

Yes. Cleaning-in-place (CIP) is a cleaning procedure that removes contaminants such as lipids, precipitates, or denatured proteins that may remain in the packed column after regeneration. Such contamination is especially likely when working with crude materials. Regular CIP prevents the build-up of these contaminants in the packed bed, and helps to maintain the capacity, flow properties and general performance of the resin.