Expanded bed chromatography for one-step purification of mannose binding lectin from tulip bulbs using mannose immobilized on DEAE Streamline

Abstract

Expanded bed chromatography allows chromatographic separations to be performed starting from unclarified material. Aiming to purify from plants mannose binding lectins, which are valuable reagents, we evaluated expanded bed methodology using immobilized mannose as an affinity chromatography support. Ligand was immobilized on DEAE Streamline, a matrix which was commercially available but obviously primarily designed for ion-exchange chromatography in expanded bed conditions. Nevertheless through adjustments of mobile phase ionic strength, it was easy to perform true biospecific affinity chromatography on this support and a single-step purification procedure for mannose binding lectins from unclarified tulip bulbs extract was devised. The cleaning in place procedure allowed repeated use of the affinity column. This report seems to be the first article on proteins purification from plant material by expanded bed chromatography and gives another example supporting the concept that expanded bed methodology may be used with very selective affinity chromatography supports.