Lysine affinity chromatography

Figure 6.2 A Likely purification procedure for tPA produced in recombinant E. coLi cells. The heterologous product accumulates intracellularly in the form of inclusion bodies. In this particular procedure, an ultrafiltra-tion step is introduced on several occasions to concentrate the product stream, particularly prior to application to chromatographic columns. Lysine affinity chromatography (Lys-chromatography) is employed, as tPA is known to bind immobilized Lysine molecules. Adapted with permission from Bio/Technology (1993), 11, 351... 

Unspecific inhibition of ribonucleotide reduction is produced by compounds like pyridoxal phosphate, or the sulfonated anthraquinone-triazine dye, Cibacron blue. They interact, like in many enzymes, with nucleotide binding domains where pyridoxal phosphate becomes covalently linked to lysine, or in that the dye occupies the whole nucleotide fold. The latter interaction permits its use in affinity chromatography of ribonucleotide reductases Likewise, EDTA is not a specific, nor a potent inhibitor, it may, for example, act by complexation of the structure-stabilizing Mg " ions in native holoenzymes. However the iron-promoted radical regeneration process appears far more susceptible to interference from EDTA ... 

Metal-chelate affinity chromatography was introduced as a specific method for fractionation of proteins by Porath et al., in 1975. The principle of this type of chromatographic method is that certain amino acid residues, such as histidine, cysteine, lysine, tryptophan, aspartic acid, glutamic acid, or phosphorylated amino acids, which are accessible on the protein surface, can interact through non-bonding lone-pair electron coordination with some metal ions. Metal cations Cu, Ni, ... 

Isolation of anti-([8-lysine]vasopressin) antibodies by affinity chromatography... 

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