Affinity chromatography application buffer

Elution is one of the critical step for successful separation. Sample application in affinity chromatography is performed usually by injection or application in the presence of mobile phase which is prepared in appropriate pH, ionic strength and solvent composition for solute-ligand binding. This solvent is referred as application buffer [8]. In the presence of application buffer, compounds which are complementary to the affinity ligand will bind while the other solutes in the sample will tend to pass through the column as nonretained compounds. After... 

A very interesting application of affinity chromatography to the purification of halophilic enzymes was reported by Sundquist and Fahey (1988). These authors have purified the enzymes bis-y-glu-tamylcysteine reductase and dihydrolipoamide dehydrogenase from H. halohium using immobilized metal ion affinity chromatography in high-salt buffers. 

The compatibility of all these PEG-based resins with aqueous buffers allows their use for biochemical applications such as on-resin screening of chemical libraries and in the development of affinity chromatography [58-61]. All these families of PEG-based resins, except POEPS, are free of aromatic rings. This feature makes these solid supports highly suitable for a broad range of applications where such rings can react with reagents or/and jeopardize the solid-phase NMR control of the reactions [62]. 

The effect of application and elution buffers must be taken into consideration in affinity chromatography. Most application buffers in affinity chromatography are solvents that mimic the pH, ionic strength, and polarity experienced by the solute and ligand in their natural environment. These conditions give the solute its highest... 

An essentially pure extracellular glycoprotein proteinase inhibitor was isolated from the latex of green fruits of papaya by a single affinity chromatography purification step. An immobilized trypsin-Sepharose CL 4B column was prepared according to the manufacturer (Amersham Pharmacia Biotech), which provided elaborated bulletins for the preparation procedures and applications of these affinity gel columns. Latex extract was applied to the column after equilibration with 20 mM Tris-HCl, pH 8.0, containing 0.5 M NaCl and 0.05 M CaCli. The column was extensively washed with the same buffer until ultraviolet (UV) absorbance became undetectible. The bound trypsin inhibitor was eluted with 0.02 M HCl and recovered by lyophilization after dialysis against water. 

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