Elution in affinity chromatography

Biospecific adsorption, 6 396-397, 399 Biospecific binding, in microarray fabrication, 16 385 Biospecific elution, in affinity chromatography, 6 398 Biosphere, selenium occurrence in, 22 78. 

Nonsolvent bath, polymer precipitation by immersion in, 15 808-811 Nonspecific elution, in affinity chromatography, 6 398, 399 Nonstationary Poisson process, in reliability modeling, 26 989 Non-steady-state conduction, 9 105 Nonsteroidal antiinflamatory agents/drugs (NSAIDs) 21 231 for Alzheimer s disease, 2 820 for cancer prevention, 2 826 Nonsulfide collectors, 16 649 Nonsulfide flotation, 16 649-650 Nonsulfide mineral flotation collectors used in, 16 648-649t modifiers used in, 16 650, 651t Nonsulfide ores, 16 598, 624... 

Hage, D.S. Xuan, H. Nelson, M.A. Apphcation and elution in affinity chromatography. In Handbook of Affinity Chromatography, 2nd Ed. Hage, D.S., Ed. Taylor Francis New York, 2006 Chapter 4. 

Hage DS, Xuan H, Nelson MA. Application and elution in affinity chromatography. In D.S. Hage DS, editor. Handbook of affinity chromatography. 2nd ed. Boca Raton, FL CRC Press 2005. p. 79-97. 

The elution of [60]- and [70]fullerenes was measured in water-methanol as a function of temperature on a poly(octadecylsiloxane) phase.67 The retention was shown to be dependent on the surface tension of the stationary phase through a simple geometrical model in which the solute formed a cavity in the stationary phase. In affinity chromatography, it was demonstrated that low ligand density may be a requirement for specificity of binding.68... 

Elovich equation, 1 595 Elution, in ion exchange, 14 410—412 Elution buffer, in affinity chromatography, 6 392... 

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... 

Firer MA. Efficient elution of functional proteins in affinity chromatography. Journal of Biochemical and Biophysical Methods 2001 49 433-442. 

Molecular exclusion chromatography is based on the inability of large molecules to enter small pores in the stationary phase. Small molecules enter these pores and therefore exhibit longer elution times than large molecules. Molecular exclusion is used for separations based on size and for molecular mass determinations of macromolecules. In affinity chromatography, the stationary phase retains one particular solute in a complex mixture. After all other components have been eluted, the desired species is liberated by a change in conditions. 

In affinity chromatography, the mobile phase is a buffer. Typically, the sample is dissolved in the buffer, and thus buffers such as potassium phosphate or (2-hydroxyethyl)-l-piperazineethanesulfonic acid (HEPES) are used. Once the unretained material has been washed from the column, the analyte must be eluted there are two approaches to the removal of the analyte nonspecific and biospecific elution. 

The method of zonal elution is one of the most common techniques used in affinity chromatography to examine biological interactions. An example of this type of experiment is shown in Fig. la. In its usual form, zonal elution involves the application of a small amount of analyte (in the absence or presence of a competing agent) to a column that contains an immobilized ligand. The retention of the analyte in this case wiU depend on how strongly the... 

As an example of the steps involved in affinity chromatography, we will consider the purification of an enzyme present in a mixture of proteins, using an immobilized competitive inhibitor as an affinity ligand. The first step after column preparation is the application of the sample during this step, the enzyme that binds to the inhibitor is retained, or adsorbed on the column. This is followed by a rinse step, whereby all nonbinding species are removed. The third step involves elution, and this step... 

Numbers in the table are molar concentrations of KCl at which subfragments were eluted from affinity chromatography columns upon increasing the concentration stepwise. Urea means that the subfragments were not eluted by a solution containing 1.0 M KCl, but were eluted with 6 Af urea (Nakamura et ai, 1981 Ohtsuki et at., 1981 Tanokura et al, 1983 Onoyama and Ohtsuki, 1986). 

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