A biopolymer-based aptasensor for the determination of patulin and the method for the development of this aptasensor
Patent Information
- Authority / Receiving Office
- EP · EP
- Patent Type
- Applications
- Current Assignee / Owner
- EGE UNIVERSITESI IDARI & MALI ISLERDAIRE BSK
- Filing Date
- 2024-04-05
- Publication Date
- 2026-05-13
AI Technical Summary
Current aptasensor designs for patulin determination are expensive, require large sample volumes, and have time-consuming modification processes, often relying on carbon-based electrodes that are not optimized for electrochemical impedance spectroscopy, limiting precision and selectivity.
Development of a biopolymer-based aptasensor using pencil graphite electrodes modified with biopolymers, where the patulin-specific aptamer sequence is immobilized, allowing for precise patulin determination through electrochemical impedance spectroscopy, enabling miniaturization and portability for field applications.
The biopolymer-based aptasensor provides precise and selective patulin determination, facilitating bedside and field monitoring of food safety and quality, with potential adaptability for various biomolecule analyses, and ease of large-scale production due to its biocompatibility and disposability.
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Abstract
Description
[0001] A BIOPOLYMER-BASED APTASENSOR FOR THE DETERMINATION OF PATULIN AND THE METHOD FOR THE DEVELOPMENT OF THIS APTASENSOR
[0002] Field of the Invention
[0003] The present invention involves the development of biopolymer-based aptasensor and the implementation of (bio)molecule analyses in a stable, inexpensive, easy-to-modify, precise and practical way, compared to the studies in the state of the art.
[0004] State of the Art of the Invention (Prior Art)
[0005] In state-of-the-art aptasensor designs, the electrode surface is modified with various carbon-based platforms, biomaterials, nanomaterials and / or polymers in order to increase determination precision and selectivity. However, the platforms used for current designs can be expensive. Large amounts of sample may be required and the modification process may also be time-consuming.
[0006] In the state of the art, there are aptasensor designs for the determination of patulin. Carbon-based electrodes are the most frequently used electrodes in these aptasensor designs. Electrodes called pencil graphite electrodes (PGE) and made by integrating graphite tips used for writing into the electrode system are frequently used in studies within the state of the art due to their advantages such as being inexpensive, providing a large surface area, and being more practical and easy to use compared to other carbon electrodes.
[0007] There are studies in the state of the art in which the determination of patulin is carried out with aptasensors.
[0008] In the patent application document number US2017 / 0153235A1 , a patulin-specific aptamer has been developed. The aptamer sequence was immobilized on the graphene oxide surface, and then fluorescence material was added to the medium to bind the fluorescence material to the aptamer sequence. Different concentrations of patulin were added to the medium and the differences in the fluorescence signal were analyzed. The study differs from the invention in that the determination of patulin is carried out based on optical methods.
[0009] In the patent application document number CN104593374A, a patulin-specific oligonucleotide aptamer has been developed. A single-chain patulin-specific oligonucleotide aptamer has been developed by the graphene oxide-based SELEX method. Determination of patulin with the developed aptamer was carried out with the fluorescence technique. The study differs from the invention in that the determination of patulin is carried out based on optical methods.
[0010] In the patent application document number CN112961862A, a patulin-specific aptamer has been optimized. The 5' or 3' end of the optimized P-30 aptamer has been modified with amino, sulfhydryl, or carboxyl. Determination of patulin with this aptamer was carried out with the fluorescence technique. The study differs from the invention in that the determination of patulin is carried out based on optical methods.
[0011] In the patent application document number CN105510570A, a method for the determination of patulin has been developed. In the method based on patulin interaction with patulin-specific aptamer, determination was carried out by glucometer. The study differs from the invention in that patulin determination is carried out using glucose with a glucometer device.
[0012] In the patent application document number CN1 11398393A, an electrochemical aptamer ratio sensor has been developed for the determination of patulin. In the study, the gold electrode surface was modified with trimetal nanoflowers, and then the nucleic acid aptamer was immobilized on the surface. Patulin determination was carried out depending on the change in the signal ratio of electroactive substances. The electrode used in the study differs from the invention in terms of the material used in modifying the electrode surface and the method by which the determination of patulin is carried out.
[0013] In the state of the art, there are aptasensor designs for the determination of patulin. Carbon-based electrodes are the most frequently used electrodes in these aptasensor designs. Electrodes called pencil graphite electrodes (PGE) and made by integrating graphite tips used for writing into the electrode system have been used since the early 2000s in studies within the state of the art due to their advantages such as being inexpensive, providing a large surface area, and being more practical and easy to use compared to other carbon electrodes.
[0014] There are studies in the state of the art in which the determination of patulin is carried out with aptasensors.
[0015] In the patent application document number US2017 / 0153235A1 , a patulin-specific aptamer has been developed. The aptamer sequence was immobilized on the graphene oxide surface, and then fluorescence material was added to the medium to bind the fluorescence material to the aptamer sequence. Different concentrations of patulin were added to the medium and the differences in the fluorescence signal were analyzed.
[0016] In the patent application document number CN104593374A, a patulin-specific oligonucleotide aptamer has been developed. A single-chain patulin-specific oligonucleotide aptamer has been developed by the graphene oxide-based SELEX method. Determination of patulin with the developed aptamer was carried out with the fluorescence technique.
[0017] In the patent application document number CN112961862A, a patulin-specific aptamer has been optimized. The 5' or 3' end of the optimized P-30 aptamer has been modified with amino, sulfhydryl, or carboxyl. Determination of patulin with this aptamer was carried out with the fluorescence technique.
[0018] In the patent application document number CN105510570A, a method for the determination of patulin has been developed. In the method based on patulin interaction with patulin-specific aptamer, determination was carried out by glucometer.
[0019] In the patent application document number CN1 11398393A, an electrochemical aptamer ratio sensor has been developed for the determination of patulin. In the study, the gold electrode surface was modified with trimetal nanoflowers, and then the nucleic acid aptamer was immobilized on the surface. Patulin determination was carried out depending on the change in the signal ratio of electroactive substances.
[0020] Brief Description and Object of the Invention This invention involves the modification of carbon-based electrodes with biopolymer and the aptamer-based determination of patulin. In the aptasensor design developed within the scope of the invention, pencil graphite electrodes (PGE) are preferably used. Although there are studies on the development of sensors based on electrodes modified with different nanomaterials and biomaterials in the state of the art, no studies have been found on the development of aptasensors involving the determination of patulin by electrochemical impedance spectroscopy technique using biopolymer-modified pencil graphite electrode.
[0021] This invention involves the biopolymer-based aptasensor, the development of this aptasensor, and the method for the determination of patulin. Carbon-based electrodes are used as the aptasensor platform, preferably pencil graphite electrodes are used. This sensor surface is modified with biopolymer. The patulin-specific aptamer sequence was immobilized onto the surface of the biopolymer-modified electrode. The interaction of patulin with patulin-specific aptamer was analyzed by electrochemical impedance spectroscopy technique. In the literature, there has been no study on the development of aptasensor involving the determination of patulin by electrochemical impedance spectroscopy technique using a biopolvmer-modified disposable graphite electrode. With the developed aptasensor, patulin determination is carried out more precisely and selectively.
[0022] Detailed Description of the Invention a) Electrochemical activation of carbon-based electrode, b) Chemical activation of carbon-based electrode, c) Modification of the electrode with a biopolymer solution, d) Immobilization of target analyte-specific aptamer sequence onto the electrode surface, e) Incubation of the target analyte, mycotoxin, on the aptasensor surface
[0023] Taking measurements by electrochemical impedance spectroscopy (EIS) technique
[0024] The aptasensor system developed within the scope of the invention can be miniaturized and compacted, making it smaller. The developed aptasensor system can be integrated into a portable impedance meter device and adapted for the impedimetric determination of patulin mycotoxin. With this feature, it can be used in bedside analyses and in the field to monitor food safety and quality. Since the developed aptasensor system is easy to use and disposable, it can be easily adapted to large-scale production. The developed aptasensor surface is biocompatible as it is modified with natural biopolymer. Therefore, it can be adapted to sensor systems to be developed for the determination of many different (bio)molecules, for use in health, food and environmental fields.
Claims
CLAIMS1 . An impedimetric-based method for detecting an analyte in a test sample in the diagnosis of patulin mycotoxin, characterized in that it comprises the process steps of: a) Electrochemical activation of carbon-based electrode, b) Chemical activation of carbon-based electrode, c) Modification of the electrode with a biopolymer solution, d) Immobilization of target analyte-specific aptamer sequence onto the electrode surface, e) Incubation of the target analyte, mycotoxin, on the aptasensor surface f) Taking measurements by electrochemical impedance spectroscopy (EIS) technique.
2. The method according to Claim 1 , characterized in that said target analyte is the patulin mycotoxin and the analyte-specific aptamer sequence is the patulin-specific aptamer sequence.
3. The method according to Claim 1 , characterized in that the electrode is a carbonbased electrode.
4. The method according to Claim 3, characterized in that the carbon-based electrode is pencil graphite electrode.
5. The method according to Claim 4, characterized in that the carbon-based electrode is screen-printed electrode, glassy carbon electrode or carbon paste electrode, etc.
6. The method according to Claim 1 , characterized in that said electrode is modified with a biopolymer material.
7. The method according to Claim 1 , characterized in that the test sample is serum or urine.
8. The method according to Claim 1 or 7, characterized in that the test sample is 30-9. The method according to Claim 1 , characterized in that the electrode is disposable or comprises a multi-electrode system.
10. An aptasensor, characterized in that it works with the method of Claim 1 .11 . The aptasensor according to Claim 10, characterized in that it can be integrated into a portable impedance meter device and adapted for the impedimetric determination of patulin mycotoxin.