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Preparation of a Monolithic Capillary Column With Immobilized Alpha-Mannose for Affinity Chromatography of Lectins

Preparation of a Monolithic Capillary Column With Immobilized Alpha-Mannose for Affinity Chromatography of Lectins

K Kishore R Tetala, Bo Chen, Gerben M Visser, Audrius Maruska, Olga Kornysova, Teris A van Beek, Ernst J R Sudhölter

J Biochem Biophys Methods. 2007 Feb 23;70(1):63-9.

PMID: 17112595

Abstract:

A simple method for the preparation of an affinity monolithic (also called continuous bed) capillary column for alpha-mannose-specific lectins is described. 2-Hydroxyethyl methacrylate in combination with (+)-N,N -diallyltartardiamide (DATD) and piperazine diacrylamide (PDA, 1,4-bisacryloyl-piperazine) as crosslinkers, were used as monomers for the monolith. After oxidation of DATD with periodate, alpha-mannose with spacer was bound to the aldehyde groups of the polymeric skeleton via reductive amination to form an affinity column for the separation, enrichment or binding studies of mannose-specific lectins. The permeability of the column was excellent. The porosity of the monolith was investigated by scanning electron microscope (SEM) and inverse size exclusion chromatography (ISEC). The affinity of the monolith was evaluated by frontal analysis (FA) and fluorescence microscopy (FM) using fluorescently labeled concanavalin (Con A). Frontal affinity chromatography showed a specific interaction of two different lectins with the alpha-mannose-modified monolith. According to FM the affinity sites were evenly distributed over the monolithic bed.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP6342172-B	Piperazine diacrylamide		6342-17-2	Price

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