G Size-Exclusion Chromatography

Wu, C. S., Senak, L., and Malawer, E. G., Size exclusion chromatography of poly(methyl vinyl ether-co-maleic anhydride) (PMVEMA). I. The chromatographic method, J. Liq.Chromatogr., 12, 2901, 1989. 

Mourey, T. H. and Bryan, T. G., Size-exclusion chromatography using mixed-bed columns with dimethylformamide at near-ambient conditions comparison of pStyragel HT Linear and PL Gel mixed-bed columns, /. Liquid Chromatogr., 14, 719, 1991. 

The architecture of hypeibranched polymers and dendrimers is connected with difficulties in determining molar mass. Many of the common characterization techniques—e.g. size exclusion chromatography (SEC)—used for polymers are relative methods where polymer standards of known molar mass and dispersity are needed for calibration. Highly branched polymers exhibit a different relationship between molar mass and hydrodynamic radius than their linear counterparts. 

Separations that are achieved on the basis of the size of the molecules (e.g. size exclusion chromatography) are not dealt with in this book. Such separations are not selective, and hence there is no selectivity to be optimized. 

From the great variety of methods for the determination of protein binding three separation methods, equilibrium dialysis (ED), ultrafiltration (UF), and ultracentrifugation (UC) and a non-conventional method with the binding to immobilized proteins has been chosen. The first methods are undoubtedly the most widely used because of their simplicity and general applicability to many different systems. Other methods e.g. size exclusion chromatography, capillary electrophoresis, or spectroscopic methods have been not described. Oravcova et al. (1996) gives a comprehensive review and comparison for these applications. 

Mori, S. Barth, H.G. Size Exclusion Chromatography Springer-Verlag Berlin, 1999. 

A significant fraction of biomolecules display natural biological affinity for certain other species, e.g. immuno-ligands, enzyme substrates, hormones. These properties can be exploited in an affinity separation process to recover and purify biomolecules in a more effective way (i.e. with higher yield and higher resolution) than can be achieved with more conventional means of purification (e.g. size exclusion chromatography). 

Prougenes, P. Berek, D. Meira, G. Size exclusion chromatography of polymers with molar mass detection. Computer simulation study on instrumental broadening biases and proposed correction method. Polymer 1998, 40, 117-124. 

Senak, L. Wu, C.S. Malawer, E.G. Size Exclusion Chromatography of Poly (vinylpyrrolidone) II. Absolute Molecular Weight Distribution by SEC/LALLS and SEC with Universal Calibration. J. Liq. Chromatogr. Relat. Technol. 1987,10 (6), 1127. 

Preston, K.R. Stevenson, S.G. Size exclusion chromatography and flow field-flow fl"actionation of wheat proteins. 

Unfortunately, because of the lability of the equilibrium in the interactions of the proteins and low molecular mass complexes indicated in fig. 1, it is difficult to determine the exact chemical speciation in such systems experimentally. Separation of fractions by mass, using, e.g., size-exclusion chromatography, upsets the equilibria under study so that results must be treated with great caution. As a result of this experimental uncertainty, computer-assisted methods have been developed which allow prediction of the chemical speciation pertaining to the low molecular mass fraction (May 1980, May et al. 1977, May and Williams 1977, Duffield and Williams 1989b). 

Figure 13 Schematic representation of (a) the four-capillary viscosity detector (reprinted from Pasch, H. Trathnigg, B. HPLC of Polymers, Springer Berlin-Heidelberg-New York, 1998 with kind permission of Springer Science+Business Media) and (b) universal calibration cunte (reprinted from Mori, S. Barth, H. G. Size Exclusion Chromatography, Springer Berlin-Heidelberg-New York, 1999, with kind permission of Springer Science+Business Media).

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