Other

University System of Maryland

Vincent L Feliccia

Richard B Thompson

Badri P Maliwal

Daniel Elbaum

Marcia W Patchan

David W Christianson

Zhengfang Ge

Carol A Fierke

The invention relates to compositions and kits for homogeneous fluorescence polarization (anisotropy) assays for detecting and quantifying metal ions in solution. Metal-dependent binding of a fluorescent ligand to an unlabeled macromolecule effects a measurable change in anisotropy as will the binding of metal ions to a fluorescent labeled macromolecule. Binding of the fluorescent ligand to the unlabeled macromolecule is metal dependent with the change in anisotropy being proportional to the concentration of bound metal ions. Conversely, if the fluorescent label is first conjugated to a macromolecule and the macromolecule is subsequently stripped of metal ion, it may then be used to signal binding of metal ions. The covalently bound fluorescent label exhibits changes in anisotropy proportional to the concentration of bound metal ions. Kits comprise a fluorescent molecule and a macromolecule.

Determination of metal ions in solution by photoluminescence anisotropy

Homogeneous fluorescence polarization (anisotropy) assays for detecting and quantifying metal ions in solution, based the metal-dependent binding of a fluorescent ligand to an unlabeled macromolecule, or the binding of a metal ion to a fluorescent labeled macromolecule. The metal-dependent binding of a fluorescent ligand to an unlabeled macromolecule (metallo-macromolecule) effects a measurable change in anisotropy as will the binding of metal ions to a fluorescent labeled macromolecule. Binding of the fluorescent ligand to the unlabeled macromolecule is metal dependent with the change in anisotropy being proportional to the concentration of bound metal ions. No binding of the fluorescent ligand to the macromolecule occurs in the absence of metal ions. Conversely, if the fluorescent label is first conjugated to a metallo-macromolecule and the metallo-macromolecule is subsequently stripped of its metal ion, it may then be used to transduce the binding of metal ions. Transduction is provided wherein the covalently bound fluorescent label exhibits changes in anisotropy proportional to the concentration of bound metal ions. In all methods, the change in anisotropy may be simply related to the metal ion concentration of the test solution.

The invention described in detail herein relates to the detection, determination, and quantitation of certain ions and small molecules in solution. The invention specifically relates to improvements in the area of photoluminescent sensors for use in a detection scheme involving the alteration of a photoluminescent label or moiety attached to or associated with an analyte binding macromolecule. One may use the changes in photoluminescence lifetime, changes in ratios of photoluminescence intensity or changes in photoluminescence polarization (anisotropy) to determine the analyte. The photoluminescence change measured correlates to the concentration of the ion or molecule in solution.

Photoluminescent sensors of chemical analytes

Growing community of inventors

Arnold, MD, United States of America

Vincent L Feliccia