Displacement chromatography protein

Torres, A. R., Krueger, G. G., and Peterson, E. A., Purification of Gc-2 globulin from human serum by displacement chromatography a model for the isolation of marker proteins identified by two dimensional electrophoresis, Anal. Biochem., 144, 469, 1985. 

Gerstner, J. A. and Cramer, S. M., Cation-exchange displacement chromatography of proteins with protamine displacers effect of salt-induced gradients, Biotechnol. Prog., 8, 540, 1992. 

Narahari, C.R., Strong, J.C., and Frey, D.D., Displacement chromatography of proteins using a self-sharpening pH front formed by adsorbed buffering species as the displacer, /. Chromatogr. A, 825, 115, 1998. 

Kundu, A., Shikla, A. A., Barnthouse, K. A., Moore, J., and Cramer, S. M., Displacement chromatography of proteins using sucrose octasulfate, BioP-harm., 64, May 1997. 

Dendrimers are being used as host molecules, catalysts, self-assembling nanostructures analogs of proteins, enzymes, and viruses and in analytical applications including in ion-exchange displacement chromatography and electrokinetic chromatography. 

Displacement Chromatography in the Separation and Characterization of Proteins and Peptides... 

As outlined above displacement chromatography may find its most important uses in the analytical area. The ability to enrich trace levels of components is ideally suited to the proteomics where more powerful tools are desperately needed to address the vast concentration ranges present in order to identify trace components. The technique also offers a way to isolate large quantities of protein variants which is important for the identification and characterization of minor product-related impurities commonly associated with therapeutic proteins. 

New concepts presented in this edition include monolithic columns, bonded stationary phases, micro-HPLC, two-dimensional comprehensive liquid chromatography, gradient elution mode, and capillary electromigration techniques. The book also discusses LC-MS interfaces, nonlinear chromatography, displacement chromatography of peptides and proteins, field-flow fractionation, retention models for ions, and polymer HPLC. 

While much has been learned about the role and selection of the operation parameters in displacement chrcmatography (60-66), little is known yet about the rules of displacer selection and the means available to control the selectivity of the separation. The paucity of well characterized displacers and the lack of knowledge of the solute adsorption isotherms hinder most strongly the wider acceptance and use of displacement chromatography. In most cases, displacer selection is still done by trial-and-error. In the majority of modem displacement chromatographic publications a reversed-phase system was used to separate small polar molecules, antibiotics, oligopeptides and small proteins (52-66). 

A. Purification of an Antigenic Vaccine Protein by Selective Displacement Chromatography... 

TABLE I High Molecular Weight Displacers Employed for the Ion-Exchange Displacement Chromatography of Proteins... 

Displacement chromatography has been recently employed for the purification of proteins from complex industrial mixtures.39,50 These industrial examples will be described in greater detail in Section VI. 

While, displacement chromatography of proteins has, for the most part, been carried out on ion-exchange stationary phases, several investigations have been carried out on stationary phases involving other modes of adsorption. 

FIGURE 5 Displacement chromatography of proteins on IMAC using imidazole as the displacer. Column SO X 5 mm i.d. Cu2+ charged metal chelate Sepharose (10 /urn) mobile phase I M NaCI, 25 mM phosphate buffer, pH 7.0 feed l.8mL of 0.89 mM ribonuclease A and 0.69 mM myoglobin in the carrier displacer 10 mM imidazole in the carrier flow rate 0.1 mL/min fraction size 100 /cL fractions. (Vunnum et a/.58)... 

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