Affinity chromatography preparative applications

Hearn, M.T.W., Smith, P.K., Mallia, A.K., and Hermanson, G.T. (1983) Preparative and analytical applications of CDI-mediated affinity chromatography. In Affinity Chromatography and Biological Recognition, pp. 191-196. Academic Press, New York. 

A third area of development in carbohydrate l.c. analyses is in the combined techniques (see Section IV,3) and other methods that provide qualitative, as well as quantitative, information about sample constituents, such as high-performance liquid affinity chromatography. The use of specific lectin- and monoclonal antibody-based, stationary phases for analytical and preparative applications is now being considered. The basic concepts of these techniques have been reviewed - and their applications to carbohydrates have been discussed. 

J. D. Protein-doped monolithic silica columns for capillary liquid chromatography prepared by the sol-gel method applications to frontal affinity chromatography. Anal Chem... 

A3, H2, S16) In general, immunological procedures have been employed for assessment of the purity of the various lipoprotein preparations. At this time, however, the availability of specific antisera may permit the application of immunological procedures in preparative work, particularly by the technique of affinity chromatography. 

Elution is one of the critical step for successful separation. Sample application in affinity chromatography is performed usually by injection or application in the presence of mobile phase which is prepared in appropriate pH, ionic strength and solvent composition for solute-ligand binding. This solvent is referred as application buffer [8]. In the presence of application buffer, compounds which are complementary to the affinity ligand will bind while the other solutes in the sample will tend to pass through the column as nonretained compounds. After... 

Successful application of affinity chromatography requires careful design of experimental conditions. The essential components, which are outlined below, are (1) creation and preparation of a stationary matrix with immobilized ligand and (2) design of column development and eluting conditions. 

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. 

Given below are examples of preparative and large-scale applications of dye-ligand chromatography which indicate that this type of affinity chromatography may be anticipated as an industrial separation technique in near the future. 

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