Recent developments in quantitative affinity chromatography

Use of a Biosensor with Surface Plasmon Resonance Detection for the Determination of Binding Constants: Measurement of Interleukin-6 Binding to the Soluble Interleukin-6 Receptor

“…The equilibrium constant for the interaction between IL-6 and sIL-6R in solution has been obtained from competitive binding studies in which the receptor solution flowing across the sensor plate was supplemented with known total concentrations of IL-6, thereby decreasing the free concentration of receptor available for interaction with the immobilized IL-6 on the sensor plate, and hence decreasing Rp. The results have been analyzed in terms of eq 3, but with Kax replaced by X^, a constitutive binding constant related to Kax by the expression (Winzor, 1992;Winzor & Jackson, 1993):…”

“…Fundamental to these studies is a quantitative understanding of the protein-protein interactions responsible for signal transduction and high-affinity binding. Here we explore application of the existing theoretical expressions for quantitative affinity chromatography (Winzor, 1992) to the characterization of the sIL-6R/IL-6 interaction by means of the BIAcore instrument.…”

Equilibrium Constants for Nonspecific Binding of Proteins to DNA May Be Obtained by Quantitative Zonal DNA Affinity Chromatography

“…The theory to obtain equilibit can be applied, is that collection of data and its analyrium constants for protein-protein or protein-small sis are simple. To analyze the zonal data, one needs ligand interactions has been established for zonal chroonly to characterize the elution time under both nonmatography experiments, in which a small band of probinding and binding conditions (3,(12)(13)(14)(15). tein is loaded to the affinity column, and for continuous elution or frontal chromatography, in which a large MATERIALS AND METHODS Proteins and DNA.…”

Insulin-like Growth Factor-binding Protein 5 Complexes with the Acid-labile Subunit

“…The broad-peaked distribution of the IGF tracer across fractions, which is seen particularly in the presence of the 6-6-5 chimera, may indicate some dissociation of previously formed complexes during fast protein liquid chromatography and a dynamic equilibrium between ternary complexes, binary complexes, and free IGF tracer. Such dissociation is well recognized in complex formation studies using fast protein liquid chromatography (19).…”