Lectins purification

Baues, R.J., and Cray, G.R. (1977) Lectin purification on affinity columns containing reductively ami-nated disaccharides./. Biol. Chem. 252, 57. 

General affinity chromatographic method for lectin purification... 

Crude chloroform-methanol-water (30 60 8, v/v) extracts of immunostainedTLC bands were analyzed without further purification by nanoelectrospray low-energy mass spectrometry. The authors showed that this effective PLC/MS-joined procedure offers a wide range of applications for any carbohydrate-binding agents such as bacterial toxins, plant lectins, and others. Phenyl-boronic acid (PBA) immobilized on stationary support phases can be put to similar applications. This technology, named boronate affinity chromatography (BAC), consists of a chemical reaction of 1,2- and 1,3-diols with the bonded-phase PBA to form a stable... 

R. Lotan, E. Skutelsky, D. Danon, and N. Sharon, The purification, composition, and specificty of the anti-T lectin from peanut (Arachis hypogaea), J. Biol. Chem., 250 (1975) 8518-8523. 

Table 6.4 Some lectins commonly used in immobilized format for the purification of glycoproteins. The sugar specificity is listed, as are the free sugars used to elute the bound glycoprotein...

Crude protein sources containing one glycoprotein usually contain multiple glycoproteins. In most such instances, lectin-based affinity systems will result in the co-purification of several such glycoproteins. 

Protein A, produced by Staphylococcus aureus, binds IgG. It is used extensively in antibody purification protocols Lectins are a group of proteins capable of binding carbohydrates. 

Lectin affinity chromatography may be used to purify glycoproteins Immobilized antibodies may be used as affinity absorbants for the antigen that stimulated their production (e.g. purification of factor VIII using immobilized anti-factor VIII antibodies)... 

Sephacryl (allyldextran cross-linked by N,N -methylenebisacrylamide) are the two types of dextran gel used in separation. Sephadex is mainly used as a glucose pol)rmer and employed for purification of many molecules such as lectins from Helix pomatia and Viciafaba, exoamylase from Pseudomonas stutzeri [14]. 

Affinity adsorbents having carbohydrate ligands have been used for the isolation and purification of many types of macromolecular substances. To illustrate, several types of antibodies, enzymes, lectins, and myeloma proteins have been obtained in highly purified form. All of the substances that have been purified by this procedure exhibit specificity for a particular carbohydrate moiety. It is the purpose of this article to assemble information on the methods for preparing affinity adsorbents having carbohydrate ligands, and to illustrate the use of these adsorbents for the purification of representative, macromolecular substances. 

In the purification of lectins, special adsorbents have been employed. The adsorbents have been prepared by attaching specific, carbohydrate ligands to activated supports.41-106,157 An affinity adsorbent prepared from chitobiose and (2-aminoethyl)-Bio-Gel was used to purify the 2-acetamido-2-deoxy-D-glucose-binding lectin from wheat germ. The data for the purification of this lectin are shown in Fig. 6. 

Another example of the preparation of a special adsorbent is the attachment of L-fucose to agarose activated with divinyl sulfone. This affinity adsorbent was prepared, and used in the purification of two different types of lectin.20 The results on the purification of one lectin on this adsorbent are shown in Fig. 7. 

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