Process for identification of compounds for modulating the activity of sodium/calcium exchange transporter
AssigneeSanofi-Aventis Deutschland GmbH
InventorsHenning Vollert, Sven Geibel, Bela Kelety, Wolfgang Doerner, Andreas Haase, Natalie Watzke, Kerstin Diekert
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Abstract
The invention refers to a cell free assay for determining the activity of a Na+/Ca2+ exchanger (NCX) protein by means of a cell free electrophysiological sensor chip, a kit of parts comprising the sensor chip with a NCX protein as well as the manufacturing and use of the kit of parts.
Claims27
An in vitro assay for determining the activity of an NCX protein wherein a sensor chip comprising said NCX protein is treated stepwise consecutively with a washing, non-activating and activating solution and the current is measured when changing from non-activating to activating treatment;
- said sensor chip comprising a body of borosilicate glass or wherein said sensor chip is covered by a mercaptane layer and has one or several insulating layers.
An assay for identifying of a compound that modulates activity of a NCX protein wherein
- a] a sensor chip comprising said NCX protein is provided, wherein the sensor chip comprises a body of borofloate-glass that carries gold structures or the sensor chip is covered by a mercaptane layer and wherein the sensor chip has one or several insulating layers;
- b] a washing solution, a non-activating solution and an activating solution are provided;
- c] a washing solution, a non-activating solution and an activating solution are provided which each of these three solutions contains a chemical compound at the same concentration in all of the three solutions;
- d] said sensor chip from a] is treated stepwise consecutively by said washing solution, said non-activating solution, said activating solution, said non-activating solution and said washing solution from b];
- e] a current is determined when changing from said non-activating to said activating solution in d];
- f] said sensor chip from e] is treated stepwise consecutively by said washing solution, said non-activating solution, said activating solution, said non-activating solution and said washing solution from c];
- g] a current is determined when replacing the non-activating by the activating solution in f];
- thereby proving modulation of the activity of the NCX protein in case the current from e] is of different strength as the current from g].
A kit of parts comprising:
- a] a sensor chip comprising NCX protein, wherein the sensor chip comprises a body of borofloate-glass that carries gold structures or the sensor chip is covered by a mercaptane layer and harboring one or several insulating layers;
- b] a washing solution;
- c] a non-activating solution;
- and
- d] an activating solution.
An assay for determining activity of NCX protein wherein a sensor chip containing a body of boro-fluoroate-glass that carries gold structures or a sensor chip being covered by a mercaptane layer and having one or several insulating layers;
- said sensor chip comprising adsorbed NCX protein containing membranes, is exposed to a rapid change in the concentration of at least one NCX substrate.
Dependencies are inferred from the claim text.