Affinity chromatography mechanism

Affinity Chromatography. This technique involves the use of a bioselective stationary phase placed in contact with the material to be purified, the ligate. Because of its rather selective interaction, sometimes called a lock-and-key mechanism, this method is more selective than other lc systems based on differential solubiHty. Affinity chromatography is sometimes called bioselective adsorption. 

Affinity chromatography is based on the lock-and-key mechanism prevalent in biological systems. The retention mechanism is very specific, but the technique is more time-consuming and more expensive than those employing other retention mechanisms. 

The mechanisms described above form the basis for the chromatographic modes described in Chapter 2, namely, normal-phase, reversed-phase, size-exclusion, ion-exchange, and affinity chromatographies. However, other modes that are variations of those mentioned above, such as hydrophobic-interaction chromatography (HIC), chiral, ion-exclusion, and ion-pair chromatographies are also used and will be mentioned. 

The basis for selectivity in affinity chromatography is the use of immobilized biochemicals, known as affinity ligands, that are covalently attached to a support matrix, as illustrated in Figure 2.17. The primary criteria that govern the suitability of a support matrix for affinity chromatography include (1) the mechanical and flow properties of the matrix, (2) the ease of covalent coupling of the ligand to the matrix, and (3) the stability of the... 

The ultimate combination of HPLC and AC is effectuated in High Performance Affinity Chromatography (HPAC).47 The development of this hybrid technique was highly assisted by the use of modified silica. Traditional polysaccharide supports may not be used for HPAC, because they lack mechanical stability to withstand the high pressure drops, inherent to this method. Modified silica beads are well suited. These may be coated with active groups as in normal AC applications. Additionally, if the separation requires the use of an organic stationary phase, the silica beads are modified with a silane or polymer with subsequent deposition of polysaccharides such as dextrans, agarose or cellulose.50... 

Arve and Liapis [34] suggest estimating the parameters characterizing the intraparticle diffusion and the adsorption-desorption step mechanisms of affinity chromatography from the experimental data obtained in a batch system. The numerical simulations of the chromatographic process will use the values of the parameters of the adsorption isotherm and those of the effective pore diffusion as determined from stirred tank experiments together with the film mass transfer coefficients calculated from chemical engineering expressions found in the literature. 

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