Affinity chromatography support materials

Support coated open tubular (SCOT) columns, 4 615 6 379 Supported liquid membranes, 16 28 Support material, in fluidized-bed encapsulation, 11 540 affinity chromatography, 6 392-393 chromatography, 6 375 gas chromatography, 6 375 Supported metals... 

For successful separation in affinity chromatography, the important parameter is that solute of interest should be bound firmly and specifically while leaving all other molecules. This requires that the support within the column contain an affinity ligand that is capable of forming a suitably strong complex with the solute of interest [8]. The other important property is that the, support material must be biologically and chemically inert to avoid... 

Porous supports like agarose, pol3mrethacrylate, or silica beads are generally used in current applications of affinity chromatography. However, in the past several years other types of supports have also become available commercially. Many of these newer materials have properties that give them superior performance in certain applications. Materials that fall in this category include nonporous supports, membranes, flow-through beads, continuous beds and expanded-bed particles. 

Initially, the antibodies should be purified prior to prepare the immunoaffinity column. Precipitation with ammonium sulfate, ion-exchange chromatography, gel filtration chraoma-tography or affinity chromatography may be employed with the aim of antibody purification. Activated beads which are coated with bacterial proteins A or G may be used as the support material. Some parameters may be changed for the elution of the sample solution for example the ionic conditions of mobile phase may be changed or chaotropic buffers may be used [11]. 

Polysaccharides that have been modified chemically, or altered physically, have been used as adsorbents for affinity chromatography. The modification of the structure of polysaccharides has been achieved by introducing cross-linkages between the chains of the polymer and bifunctional reagents. The alteration of the properties of polysaccharides by physical means can be effected by embedding the polysaccharide in a network of the support material. The molecular in-... 

Affinity chromatography has become one of the standard techniques for the separation and purification of enzymes. It is based on the idea of utilizing the specific interaction between an enzyme and an immobilized substrate or inhibitor. The substrate is chemically bound to an inert support material, and when a sample is added to the column the enzyme binds to it to an extent depending on the strength of the interaction between the two. Other enzymes and proteins which do not interact with the substrate pass through the system with little or no retention. Removal of the enzyme from the stationary support is accomplished by changing the eluent so as to alter the enzyme-substrate interaction. The... 

GPC is a further special form of liquid chromatography. The separation column is packed with porous, polymer gels (e.g. polystyrene gel) as stationary phase. The particle size of the packing material and the size distribution of the pores are well defined and uniform. In GPC molecules are separated according to their effective size in solution, i.e., their hydrodynamic volume, and not according to their affinity for the support material. 

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