Size exclusion chromatography process

Column Si. Size-exclusion chromatography columns are generally the largest column on a process scale. Separation is based strictly on diffusion rates of the molecules inside the gel particles. No proteins or other solutes are adsorbed or otherwise retained owing to adsorption, thus, significant dilution of the sample of volume can occur, particularly for small sample volumes. The volumetric capacity of this type of chromatography is determined by the concentration of the proteins for a given volume of the feed placed on the column. 

The total stationary-phase volume required to process a given feed stream is proportional to the inlet concentration and volume of the feed. For example, for a typical inlet concentration of protein of 10 g/L, in a 100 L volume of feed, a column volume of at least 100 L is needed for size-exclusion chromatography. In comparison, an ion-exchange column having an adsorption capacity of 50 g/L would only require 20 L of column volume for the same feed. 

Toyopearl HW size exclusion chromatography resins are macroporous packings for bioprocessing chromatography. They are applicable for process-scale... 

It is basically a fractionation process that depends not only on molecular size, but also on chemical composition, stereo-configuration, branching, and crosslinking. For multicomponent systems, fractionation with different ion polymolecularity, chemical heterogeneity and sequence length distribution, solubility or elution fractionation is of primary importance. Therefore, gel permeation chromatography or size exclusion chromatography is used as an important tool for the characterization of PBAs. 

Advanced computational models are also developed to understand the formation of polymer microstructure and polymer morphology. Nonuniform compositional distribution in olefin copolymers can affect the chain solubility of highly crystalline polymers. When such compositional nonuniformity is present, hydrodynamic volume distribution measured by size exclusion chromatography does not match the exact copolymer molecular weight distribution. Therefore, it is necessary to calculate the hydrodynamic volume distribution from a copolymer kinetic model and to relate it to the copolymer molecular weight distribution. The finite molecular weight moment techniques that were developed for free radical homo- and co-polymerization processes can be used for such calculations [1,14,15]. 

The two techniques differ in that HDC employs a nonporous stationary phase. Separation is affected as a result of particles of different size sampling different velocities in the interstitial spaces. Size exclusion chromatography is accomplished by superimposing a steric selection mechanism which results from the use of a porous bed. The pore sizes may vary over a wide range and the separation occurs as a result of essentially the same processes present in the gel permeation chromatography of macromolecules. 

IEC is also important in large-scale manufacturing of gene vectors such as plasmid DNA. In one process flow scheme, IEC and size exclusion chromatography (SEC) were used sequentially to purify the plasmid after lysis... 

Reynolds, J. G., and Biggs, W. R., Application of Size Exclusion Chromatography Coupled With Element-Specific Detection to the Study of Heavy Crude Oil and Residua Processing Accounts of Chemical Research, 1988. 21 pp. 319-326. 

Figure 19.10. Summary of purification processes for two human proteins synthesised in recombinant E. colt6 ). The term gel filtration is sometimes used as an alternative to size-exclusion chromatography...

This is the consequence of the traditional application of CE in the process and product monitoring of rDNA-derived biopharmaceuticals in biotechnological industries. However, related proteins, and dimer and related substances of higher molecular mass of somatropin, and aprotinin are evaluated by means of HPLC and size exclusion chromatography, respectively, by the EP. 

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