Size-exclusion chromatography specific resolution

The selection of the particle size of a column involves several compromises. One is the compromise between analysis time, resolution, and backpressure discussed in general in Chapter 3. The second is the selection of the range in the chromatogram in which we desire maximum resolution and the conditions associated with this. The third is the compromise between the desire to maximize resolution and the prevention of breakdown of the sample due to shear degradation. The second and third are specific to size-exclusion chromatography. 

Resolution in size-exclusion chromatography is a function of both the calibration curve and column performance. To normalize resolution to a molecular-wei t difference of a decade, the specific resolution has been defined as... 

Size exclusion chromatography (or gel filtration ) has a unique set of attributes which sets it apart from other methods. Its applications are limited because it has low resolution, low capacity, and is slow. However, it is the only practical separation method based on size, whidi is a relatively invariant property of immunoglobulins in contrast to charge, hydrophobidty, and other specific binding interactions. The resolution achieved by SEC may not be spectacular however separations are predictable and nearly independent of buffer conditions. so the need for method development is minimal. 

Specific protein components in food/food products can be determined by chromatography (ion-exchange chromatography, PC, TLC, GC, LC, etc.), electrophoresis, and immunology or their combination. It is also necessary to use high-resolution techniques such as reversed phase LC, ion-exchange LC, size-exclusion LC, slab electrophoresis, and capillary electrophoresis (CE). 

Following initial purification, final purification via chromatography is required. This is well known for its high resolution and purification that removes impurities and brings the product close to final specifications. There are many chromatographic techniques such as size exclusion, ion exchange, hydrophobic interactions, and affinities that can be used for final enzyme purification. Each has its limitations that must be taken into consideration before selection. 

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