A simple and convenient approach for evaluation of the parameters of ligand–receptor interaction. Receptor blocking index and its application

Abstract

A new approach for determination of the parameters for ligand-receptor interaction, which is based on so-called dilution coordinates, was developed earlier. Equations that allow evaluation of not only the affinity of ligand-receptor interaction but also of the amount of free (or occupied by corresponding ligand) receptors were suggested. The most important advantage of this approach as compared with well-known methods is the ability to determine the binding parameters for ligand-receptor interaction even for the cases in which ligand and receptor are already present in a mixture and separation of counterparts from each other is technically difficult or even impossible. Due to this reason, the proposed approach can be especially useful for studying interactions between highly-labile biological receptors and corresponding ligands as found in vivo. In the present paper I continue to consider how to determine the binding parameters for a given ligand-receptor interaction if the value of receptor blocking index is determined experimentally.

A Single D_H Gene Segment Is Sufficient for the Establishment of an Asthma Phenotype in a Murine Model of Allergic Airway Inflammation

“…These findings are further sustained by our observation that the affinity constants (K a ) of the allergen-induced IgG 1 antibodies did not differ between ΔD-DFL and WT mice. Interestingly, the K a of allergen-specific immunoglobulins in our study lay in the same order of magnitude as the K a of previously published monoclonal OVA-specific IgG 1 antibodies [20]. …”

“…In OVA-sensitized and OVA-challenged animals, K a values did not differ significantly between the two genotypes (p = 0.72) (table 1). The K a of allergen-specific immunoglobulins in our study lay in the range of one third of the K a of previously published monoclonal OVA-specific IgG 1 antibodies [20]. …”

“…Antibody affinity constants were determined by ELISA, as previously described [20]. Briefly, pooled serum was diluted by 0.1% BSA solution in PBS to give a final concentration of 2 ng/ml.…”