Method for Detecting Second Messengers with Fluorescence-Containing Sensor Proteins and Device for Carrying Out Said Method

Simple SummaryContent extracted from patent full text and abstract with AI.

This patent describes a method, device, and kit for detecting and quantifying 'second messengers'—small molecules that relay signals inside biological cells, such as cyclic AMP (cAMP)—using special fluorescent sensor proteins. The procedure involves adding the sensor protein to a container, measuring its baseline fluorescence, introducing the target molecule (e.g., cAMP), and detecting the resulting change in fluorescence. The system is designed to be highly sensitive, rapid, robust, and cost-effective, making it suitable for use outside of living cells (in cell extracts or biochemical assays). The patent also covers the preparation and storage of these sensor proteins so they can be easily used in multiwell plates for high-throughput measurements.

Use CasesContent extracted from patent full text and abstract with AI.

• High-throughput screening of drug candidates affecting cellular signaling pathways involving cAMP, cGMP, or other second messengers.
• Routine laboratory assays to quantify second messengers in cell or tissue extracts for cell biology, pharmacology, and neuroscience research.
• Automated or semi-automated diagnostic tools for research or clinical settings to monitor cell signaling changes.
• Educational kits for hands-on learning in biochemistry and molecular biology.
• Research on signal transduction, GPCR pharmacology, and second messenger pathways in basic and applied science.

BenefitsContent extracted from patent full text and abstract with AI.

• Much faster and less labor-intensive than traditional methods like HPLC or radioimmunoassays for detecting cAMP or similar molecules.
• Avoids the use of radioactive substances, making it safer and simpler for laboratories to implement.
• Sensor proteins can be prepared, stored (dried or immobilized), and rapidly reconstituted for use, improving convenience and stability.
• Compatible with widely used multiwell plate formats, enabling high-throughput and parallel analyses.
• Sensitive over a wide range (four orders of magnitude) and robust to varying buffer conditions—suitable for diverse experimental setups.
• Cost-effective due to simple assay requirements and the ability to produce large amounts of sensor protein via bacterial expression systems.
• Reduces sample preparation and incubation times dramatically (results in ~30 minutes), increasing laboratory efficiency.
• Versatile: while primarily shown for cAMP, the method can be adapted for other second messengers (e.g., cGMP, IP3) and assay types.

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Patent Abstract

The invention relates to a method for detecting second messengers, characterised by the following steps: adding a fluorescence-containing sensor protein to a container; measuring the basic fluorescence without the second messenger; adding the second messenger to the container; and measuring the change in fluorescence in order to detect the second messenger. The invention also relates to a device and a kit for carrying out said method.