Receptor–ligand binding assays: Technologies and Applications

Thermodynamic and kinetic characterization of pH-dependent interactions between bovine serum albumin and ibuprofen in 2D and 3D systems

“…Thus, the quantitative study of the binding thermodynamics and the knowledge of the kinetics of the release process are necessary [9]. Contrary to the (radio)-labelled techniques in recent years a number of "label-free" techniques have been developed to report biomolecular interactions [10][11][12]. Two-dimensional SPR is a label-free technique and capable of measuring real-time quantitative binding affinities and kinetics for proteins interacting with biomolecules using relatively small (in nanomolar range) quantities of materials and has potential to be medium-throughput [13][14][15].…”

A Fluorescent Ligand-Binding Alternative Using Tag-lite® Technology

“…Since FP does not require any receptor tagging, binding assays can be carried out on wild-type receptors or on endogenous receptors if well expressed. However, it has several drawbacks such as (1) fluorescent ligand mass should be below 5 kDa and have high affinity (below 5 nM), 2 (2) fluorescence background from chemical compounds or biological material can affect the accuracy of FP, and (3) the hydrophobic nature of some small fluorescent ligands can induce significant nonspecific binding to cell membranes, resulting in a small output signal.…”

Biomarker Quantitation: Analytical Considerations for Ligand Binding Assay Regression Curves and Quality Control Samples

“…Many of these fall into the ligand binding assay category (Sittampalam et al, 1997;Jong et al, 2005), specifically the microtiter variety. Standard EnzymeLinked Immunosorbent Assays (ELISA) have been around for decades, (Weeman and Schuurs, 1971;Engvall and Perlmann, 1971) and although they are known to have fairly narrow dynamic ranges (up to 1½ logs), these assays are simple and well established in most bioanalytical labs (Porstmann and Kiessig, 1992).…”