An organic electrochemical transistor (OECT)-based biosensor for detecting DNA interacting proteins includes: a substrate; multiple electrodes including a source, a drain, and a gate; a conducting channel connecting the source and drain; and an active bio-functionalized layer on the conducting channel. The bio-functionalized layer comprises a top layer of biotinylated-DNA (B-RF) designed to recognize and bind specific proteins. Additionally, a process for preparing these biosensors is provided.
Figure (1) organic electrochemical transistor (OECT)
Monitoring protein-DNA interactions is crucial for understanding essential biological processes like transcription, recombination, and replication. Conventional biochemical methods such as DNA footprinting and electrophoretic mobility shift assays are laborious and require extensive sample preparation. While electrochemical techniques have been successful in detecting biomarkers and small molecules, there has been limited exploration in tracking protein-DNA interactions.
- Ideal for detecting DNA binding proteins
- Simple fabrication and operates at low voltage
- Enables efficient and selective protein detection
- Modular sensor design
- Adaptable to target various proteins by altering the top functionalized layer (biotinylated-Bs)
- Capable of detecting biomolecules (Tp) in complex mixtures or crude cell lysates
- Operates directly for protein detection without additional preprocessing steps
The OECT developed in this invention details its fabrication process and operational methodology. In-house purified proteins such as Fob1 and FoxO1 are detected using this device. The OECT device is constructed with a PET/glass substrate as the base, an organic semiconductor channel (preferably PEDOT:PSS) connecting the source and drain, and a top gate electrode made from nickel alloy. The channel (PEDOT:PSS organic layer) of the OECT is then bio-functionalized for the specific detection of DNA binding proteins.
The bio-functionalization process involves: (1) Spin-coating an APTES functionalized layer on top of PEDOT:PSS, (2) Applying a biotin layer on (1) using sulfo-NHS-biotin, (3) Depositing streptavidin on layer (2) to form a biotin-streptavidin module, and (4) Creating a biotinylated DNA layer on (3), where the DNA strand facilitates the detection of proteins. Once fabricated, the OECT can directly operate with modifications, allowing subsequent protein detection through changes in channel current using a parameter analyzer instrument.
The invention is at the stage of Actual system ready for commercial deployment.
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This technology facilitates precise and efficient detection of protein-DNA interactions, crucial for understanding cellular processes and disease mechanisms. Its modular design allows for versatile application in various fields, including medical diagnostics, pharmaceutical development, and environmental monitoring.
This invention focuses on developing an OECT platform capable of selectively detecting protein-DNA interactions. Accurate detection of these biomolecular interactions is critical for understanding fundamental cellular processes. The invention presented in the work demonstrates the proof-of-concept (TRL level 3) and are validated in laboratory environment (TRL level 4) with proteins like Fob1 and FoxO1 which are of utmost importance in cell and responsible for aging. The invention also emphasizes on the fact that, the DNA strand used in the final functionalization step can be changed according to the targeted protein demonstrating the modular and multiplexing prospective of the OECT.
Geography of IP
Type of IP
202221058953
431170