Affinity chromatography adsorbent preparation

With the advent of affinity chromatography to isolate enzymes [7] it was possible to adapt the method for the isolation of antibodies [8], The purification of many types of anti-carbohydrate antibodies with specificity for carbohydrate residues of macromolecules has been achieved. The antibodies were isolated by affinity chromatography on adsorbents with proper ligands. The immunodeterminants of the antigens have been found to be mono- or disaccharide residues of the antigen. The anti-carbohydrate antibodies with unique properties are useful in a variety of medicinal and technological applications. In this article the emphasis is on polyclonal antibodies specifically the preparation, properties and uses of anti-carbohydrate antibodies. 

For every protein purification problem there is always an affinity solution, but cost and safety considerations may render these solutions impractical. As an example, antibodies are widely used for analysis, where only relatively small amounts are usually required, but their production and purification on a large scale for preparative-scale chromatography may be difficult to justify economically. In some cases, Hhybridoma technology may be able to address this problem. Even if production costs are acceptable, the immobilized antibodies may be unstable over the sequence of sample application, elution, and sanitation required for multiple use of the affinity adsorbent. For these reasons, while biological ligands (antibodies, enzymes, receptors, lectin. 

Kasai, K. Ishii, S. Affinity chromatography of trypsin and related enzymes. I. Preparation and characteristics of an affinity adsorbent containing tryptic peptides from protamine as ligands. J. Biochem. Tokyo 1975, 78, 653-662. 

A rapid procedure involving affinity chromatography on immobilized fetuin has been used to remove contaminating proteases from commercial preparations of neuraminidase. " The affinity-adsorbed enzyme was virtually free from protease activity, and the use of protease-free neuraminidase in studies of the metabolism of desialylized glycoproteins and of tumour cells was discussed. 

Sundberg, L., and Porath, J. (1974) Preparation of adsorbents for biospecific affinity chromatography. 

Many other means of preparation of adsorbents for affinity chromatography are also available.121122 For example, l,l -carbonyl-diimidazole can be used to couple a diamine to the matrix (Fig. 3-11). This reagent has the advantage that it does not depend upon the relatively unstable isourea linkages formed by Eq. 3-9 to hold the specific affinity ligands.121... 

Sundberg, L., and Porath, J. (1974) Preparation of adsorbents for biospecific affinity chromatography. I. Attachment of group containing ligands to insoluble polymers by means of bufunctional oxiranes. /. Chromatogr. 90, 87—98. 

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