Dye-ligand Chromatography

Abstract

The availability of proteins and enzymes in pure form is becoming increasingly important to genetic engineering technology, clinical diagnosis and in vivo therapy, as well as the pharmaceutical and fine chemicals industry. Biochemicals such as proteins and enzymes may be isolated directly from animal sources, cells or tissue cultures as well as native or genetically engineered microbial cells. The need for more rigorous quality criteria imposed on such products in order to satisfy current regulations has contributed to the remarkable development and expansion of affinity chromatography as a powerful separation technique (Lowe, 1979a; Scouten, 1981; Dean et al., 1985; Lowe and Clonis, 1985; Clonis, 1987; Clonis and Lowe, 1987). In principle, the technique exploits the formation of specific reversible complexes formed between the immobilised ligand and the complementary biomolecule(s) to be isolated. Depending on the ligand immobilised, affinity systems may be distinguished as those of high specificity, capable of binding usually one biomolecule, and those of moderate specificity, the so-called group-specific adsorbents, capable of binding a number of similar biomolecules. In the latter category one finds coenzymes, lectins, phenylboronates, nucleotides, polynucleotides, heparin, protein A, organomercurials and amino acids (Lowe, 1979a; Scouten, 1981; Clonis, 1982a; Chaiken et al., 1983).