Affinity chromatography mono-specific ligands

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. 

Enzyme purification was the first application of bioaffinity chromatography [23] and remains an important use of this technique. In this type of separation, ligands, such as enzyme inhibitors, coenzymes, or cofactors, are used to purify and separate enzymes [25]. For instance, in 1968 and the first report of "modem" affinity chromatography, Cuatrecasas, Wilchek, and Anfinsen employed specific enzyme inhibitors to selectively isolate enzymes [1,4]. A more recent example was the use of a support containing flavin mononucleotides for the purification of flavin adenine dinucleotide synthetase [26]. Other examples have included the use of mono-, di-, and triphosphate nucleotides for the purification of kinases and the use of nicotinamide adenine dinucleotide for the isolation of dehydrogenases [26,27]. 

The enzyme was purified 178-folds with 90-95% homogeneity and showed a specific activity of 7 umol NADH oxidised min—1 mg—1 protein by emplying ion-exchange (DEAE cellulose. Mono Q), gel filteration (Sephacryl S-300) and two affinity column chromatography. The best separation of the enzyme protein was obtained with nucleotide analogue affinity ligand reactive-red 120 agarose. The enzyme... 

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