Patent Application
20220334078
The present invention relates to the field of organic electrochemical transistor-based sensors (OECT sensor). In particular, the present invention relates to the field of sensors for biologic analytes.
Organic electrochemical transistors (OECT) have been extensively studied for the applications of sensors due to their low operating voltages, simple structure and ability to operate in aqueous environments. These features are essential for biological applications. OECTs based on several different conducting polymers have been developed for chemical and biological sensing, including humidity sensors, pH and ion sensors, biologic analyte sensors such as glucose sensor, lactose sensor, cholesterol sensor, uric acid sensor, tyrosine sensor, ascorbic acid sensor, dopamine sensor, adrenaline sensor, gallic and sialic acid sensor and cell-based biosensors.
OECT sensors are used for detection, i.e. a qualitative response to a condition, as well as measure of concentration of an analyte, i.e. for quantitative measurement. For instance, highly sensitive glucose sensor is useful in the monitoring of diabetes. OECT glucose sensors have been reported in US2012/0247976: they consist in an OECT structure whose gate is modified with a layer of Glucose Oxidase and chitosan polymer. However, such device does not detect high concentrations of glucose (e.g. hyperglycemic values) and is not suitable for in-vivo use.
OECT sensors with improved performances are thus desirable, especially for real-time monitoring concentration of biologic analytes, and in particular for in-vivo monitoring.
It is therefore an object of the present invention to provide an OECT sensor with high sensibility and robustness, and/or with a wide range of measurement of concentration adapted to in vivo detection.
The present invention relates to an organic electrochemical transistor comprising source and drain connected by a conductive channel, a gate electrode, a catalytic layer and a biologic detection layer, wherein the catalytic layer is a composite material comprising noble metal nanoparticles and a conductive matrix selected from an organic conductive polymer or a conductive allotrope of carbon, wherein the catalytic layer is in direct contact with the gate electrode, and wherein the biologic detection layer is in direct contact with the catalytic layer.
In an embodiment, noble metal nanoparticles are platinum nanoparticles.
In an embodiment, noble metal nanoparticles are grafted with thiophenol derivatives of formula (I):
In a configuration of this embodiment, R moiety of thiophenol derivatives is charged, preferably R moiety comprises a function selected from charged ternary amine, quaternary amine, sulfonate and phosphonate, more preferably thiophenol derivative is selected from 4-trimethylammonium thiophenol (IIa), 4-mercaptobenzenesulfonic acid (IIb), N-4-thiophenol-(2, 5 diamino) propenamide (IIc) and 4-mercaptobenzenephosphonic acid (IId):
In another configuration of this embodiment, R moiety of thiophenol derivatives comprises at least one aromatic group, preferably thiophenol derivative is selected from N-(4-mercaptophenyl)pyrene-1-carboxamide (IIIa) and 4-(tritylamino)benzenethiol (IIIb):
In an embodiment, the organic conductive polymer of the catalytic layer is selected from polythiophene derivatives, polypyrrole or polyaniline, preferably organic conductive polymer of the catalytic layer is poly(3,4-ethylenedioxythiophene) polystyrene sulfonate.
In an embodiment, the conductive allotrope of carbon of the catalytic layer is selected from carbon nanotubes, graphene or carbon powder.
In an embodiment, the biologic detection layer is a composite material comprising a biologic recognition element and a polymer, preferably a polymer selected from polyvinylpyridine derivatives or polysaccharides derivatives such as chitosan derivatives or cellulose acetate derivatives, more preferably a chitosan derivative.
In an embodiment, the biologic detection layer is a composite material comprising a biologic recognition element and an allotrope of carbon, preferably carbon nanotubes, graphene or carbon powder.
In an embodiment, the biologic recognition element is an enzyme, preferably Glucose oxidase or Lactate oxidase.
In a specific configuration, the organic electrochemical transistor comprises:
and
In an embodiment, the organic electrochemical transistor is totally or partially encapsulated in a membrane.
The present invention also relates to a method for detection of a biological analyte wherein a biological fluid is contacted with an organic electrochemical transistor as described above.
In the present invention, the following terms have the following meanings:
“Organic electrochemical transistor” or “OECT”: refers to a device comprising three electrodes: (2S) the source or source electrode, (2D) the drain or drain electrode, and (2G) the gate or gate electrode. In an OECT, the source and drain electrodes are connected by a conductive polymer which acts as a channel (3); and the channel and the gate electrode are separated by an electrolyte (4) which acts as gate dielectric. State of the art OECT structure is displayed in
The following detailed description will be better understood when read in conjunction with the drawings. For the purpose of illustrating, the OECT is shown in the preferred embodiments. It should be understood, however that the application is not limited to the precise arrangements, structures, features, embodiments, and aspect shown. The drawings are not drawn to scale and are not intended to limit the scope of the claims to the embodiments depicted. Accordingly, it should be understood that where features mentioned in the appended claims are followed by reference signs, such signs are included solely for the purpose of enhancing the intelligibility of the claims and are in no way limiting on the scope of the claims.
This invention relates to an organic electrochemical transistor (OECT) comprising source and drain connected by a conductive channel, a gate electrode, a catalytic layer and a biologic detection layer, wherein the catalytic layer is a composite material comprising noble metal nanoparticles and an organic conductive matrix.
Noble metal nanoparticles behave as catalytic sites and need to be accessible. However, nanoparticles are usually dispersible in aqueous solutions: they may detach from OECT leading to loss of sensibility or even failure of the device.
Noble metal nanoparticles are preferably spherical, with diameter in a range from 0.1 nm to 200 nm. According to an embodiment, noble metal nanoparticles diameter is about 1, 2, 3, 5, 7, 10, 12, 15, 20, 25, 30, 35, 40, 50, 60, 70, 85, 100, 125, 150, 175 or 200 nm.
In the invention, nanoparticles are included in an organic matrix, forming a composite material. Consequently, nanoparticles are bound to the composite material by intermolecular interactions (i.e. Van der Walls and electrostatic interactions) and/or geometrical constraints, and nanoparticle release is avoided.
In addition, matrix allows building a catalytic layer with large dimensions, at least with a thickness much larger than a layer of nanoparticles. Such thick material presents an increased contact surface for nanoparticles with sample under study, and allow to engage more nanoparticles in the device. An improved sensibility is obtained.
In an embodiment, noble metal nanoparticles are present only in catalytic layers. In other words, electrodes (source, drain and gate) and channel of OECT do not contain noble metal nanoparticles. Indeed, if noble metal nanoparticles are inside electrodes or channel, oxidation may occur in the electrode or channel material, leading to noisy signal.
In a preferred embodiment, the catalytic layer is in direct contact with gate electrode of OECT and may be on a part of the gate electrode, i.e. catalytic layer may not cover the whole surface of gate electrode. Direct contact of the catalytic layer and the gate electrode of OECT limits noise and artefacts in signal.
In a preferred embodiment, electrodes are metallic, especially gold (Au), Silver (Ag) or platinum (Pt); or conductive allotrope of carbon, especially graphene, carbon nanotubes or pure carbon particles. Electrodes are on a substrate which may be selected from various polymers, glass or semi-conductor materials. Electrodes may be deposited by ink-jetting, vacuum deposition, spin coating, spray coating or any other deposition technique.
According to an embodiment, noble metal nanoparticles are platinum nanoparticles. Platinum nanoparticles are indeed very efficient in oxydo-reduction processes, hence enabling electron transfer from an analyte to be detected into OECT.
According to an embodiment, noble metal nanoparticles are grafted with thiophenol derivatives of formula (I):
By grafted, it is meant that a bond is created between the noble metal nanoparticle and the thiophenol derivative: nanoparticle-S-C6H4-R.
By grafting, nanoparticle surface is functionalized by moiety R and compatibility with other materials is improved.
In particular, with charged R moieties, nanoparticles are easily dispersed in a charged material via electrostatic interactions. In addition, surface of such nanoparticles is easily accessible in aqueous medium. Charged R moiety may be selected from charged ternary amine, quaternary amine, sulfonate and phosphonate. Particularly suitable thiophenol derivative are 4-trimethylammonium thiophenol (IIa), 4-mercaptobenzenesulfonic acid (IIb), N-4-thiophenol-(2, 5 diamino) propenamide (IIc) and, 4-mercaptobenzenephosphonic acid (IId).
For instance, platinum nanoparticles functionalized with charged moieties are easily dispersed in PEDOT:PSS conductive polymer, yielding a very suitable composite material for invention.
Alternatively, with aromatic R moieties, nanoparticles are easily dispersed in aromatic materials via Π-Π interactions. In addition, surface of such nanoparticles is easily accessible in organic medium. Aromatic R moiety may be selected from N-(4-mercaptophenyl)pyrene-1-carboxamide (IIIa) (also known as N-4-thiophenol-pyrene-1-carboxamide) and 4-(tritylamino)benzenethiol (IIIb) (also known as 4-(1-[(triphenylmethyl)amino]) thiophenol).
For instance, platinum nanoparticles functionalized with aromatic moieties are easily dispersed in aromatic conductive polymer, yielding a very suitable composite material for invention.
According to an embodiment, the organic conductive matrix of the catalytic layer is selected from a conductive polymer and a conductive allotrope of carbon. Preferably conductive matrix of the catalytic layer is selected from polythiophene derivatives, polypyrrole, polyaniline, carbon nanotubes, graphene and carbon powder. More preferably conductive matrix of the catalytic layer is poly(3,4-ethylenedioxythiophene) polystyrene sulfonate.
According to an embodiment, the biologic detection layer is a composite material comprising a biologic recognition element and an organic matrix. In particular, the organic matrix of the biologic detection layer is selected from a polymer and an allotrope of carbon, preferably organic matrix of the biologic detection is selected from polysaccharides derivatives such as chitosan derivatives or cellulose acetate derivatives, polyvinylpyridine derivatives, carbon nanotubes, graphene and carbon powder, more preferably organic matrix of the biologic detection layer is a chitosan derivative, in particular a partially deacetylated chitin. In a preferred embodiment, biologic recognition element is an enzyme, in particular Glucose oxidase or Lactate oxidase.
In a preferred embodiment, the biologic detection layer is in direct contact with the catalytic layer and may be on a part of the catalytic layer, i.e. biologic detection layer may not cover the whole surface of catalytic layer. In a more preferred embodiment, biologic detection layer is in direct contact with the catalytic layer, which is also in direct contact with gate electrode of OECT.
The function of biologic recognition element is to react with the biologic analyte of interest, thus producing an electrochemically active compound. The later may interact with noble metal nanoparticles as catalyst to be reduced, thus generating an electric charge finally detected by OECT.
In an advantageous embodiment illustrated in
In the specific embodiment using Glucose oxidase as biologic recognition element and platinum nanoparticles in PEDOT:PSS as catalytic layer, Glucose oxidase reacts with glucose from sample and produces hydrogen peroxide H2O2. Hydrogen peroxide is oxidized in presence of platinum nanoparticles and reduces charge density in catalytic layer, yielding a variation of potential in gate electrode and a signal in OECT device.
Alternatively, enzyme used as biologic recognition element is Lactate oxidase. Lactate oxidase reacts with lactate from sample and produces pyruvate and hydrogen peroxide H2O2. Hydrogen peroxide is then oxidized in presence of platinum nanoparticles and reduces charge density in catalytic layer, yielding a variation of potential in gate electrode and a signal in OECT device
According to an embodiment, the organic electrochemical transistor is totally or partially encapsulated in a membrane. This membrane may have several functions. Membrane may limit biofouling, thus improving biocompatibility and allowing for in-vivo use. Membrane may also selectively prevent diffusion of interferent species (large molecules, cells fragments, pollutants) towards OECT, thus improving sensibility, accuracy and/or time life of OECT. Membrane may also impose a difference in concentration of the biological analyte of interest between sample in contact with membrane and electrolyte in contact with biologic detection layer: such partition may improve accuracy of measurement or improve range of measurement of OECT.
The invention also relates to a method for detection of a biological analyte wherein a biological fluid is contacted with an organic electrochemical transistor as described above. Biological analytes of particular interest are glucose and lactate. The method may be implemented in-vivo. The method may be implemented as real-time monitoring.
While various embodiments have been described and illustrated, the detailed description is not to be construed as being limited hereto. Various modifications can be made to the embodiments by those skilled in the art without departing from the true spirit and scope of the disclosure as defined by the claims.
The present invention is further illustrated by the following examples.
Preparation of Grafted Platinum Nanoparticles
Under argon using Schlenk techniques, 4-aminophenyl disulfide (496 mg, from Aldrich) is dissolved in anhydrous DMF (10 mL) under stirring. N,N-diisopropylethylamine (2.04 mL, from Aldrich) and iodomethane (2.24 mL, from Aldrich) were added and the mixture was stirred for 48 h at room temperature. The solution was filtered, and the crude product was precipitated by adding dropwise the filtrate solution to 100 mL of vigorously stirred cold diethylether. The solution was filtered and the residual solid was purified by washing it twice with boiling acetone in a reflux apparatus for 45 min. The colorless powder was dried under high vacuum leading to the desired product (432 mg).
Synthesis of Platinum Nanoparticles with a 4-(Trimethylammonium)Thiophenolate Crown
A solution of Platinum(IV) chloride (387.3 mg, from Aldrich) dissolved in hexylamine (95 mL, from Acros Organics) was prepared (Solution A). A solution of sodium borohydride (349.9 mg, from Aldrich) in a mixture of 25 mL distilled water and 25 mL methanol (from VWR) was prepared and 25 mL hexylamine were added (Solution B) Immediately after preparation, the solution B is combined with the solution A (Solution C). After 20 s, a solution of 4,4′-disulfanediylbis(N,N,N-trimethylbenzenamonium) (425 mg) in a mixture of 25 mL of hexylamine, 25 mL of methanol and 15 mL of distilled water was added to the solution C. After 3 min 30 s, 110 mL of distilled water were added, and the mixture was stirred at room temperature for 10 min. The reaction mixture was then transferred into a separation funnel and 150 mL of distilled water were added. The aqueous phase was isolated and concentrated under reduced pressure. The nanoparticles were dispersed in 30 mL ethanol (from VWR) and 4,4′-disulfanediylbis(N,N,N-trimethylbenzenamonium) (426 mg) and 15 mL distilled water were added. The reaction mixture was stirred overnight at room temperature. The reaction mixture was then centrifugated at 7000 rpm for 20 min. The supernatant was removed, and the residual powder was redispersed in a mixture of 25 mL ethanol and 25 mL distilled water then centrifugated at 7000 rpm for 20 min twice. Finally, the powder was redispersed in absolute ethanol (from VWR) and centrifugated at 7000 rpm for 20 min. The clear supernatant was removed, and the black powder was left to dry in air overnight and then 3 h under high vacuum to obtain the platinum nanoparticles (344 mg).
Diameter of nanoparticles is determined to be 30 nm.
Preparation of OECT-1
A substrate (1) of Kapton® HN (general-purpose polyimide film supplied by DuPont) is cut to appropriate dimensions and cleaned by successive washing with acetone, water and isopropanol, then dried and heated for 60 minutes at 180° C.
A mask is prepared according to geometry of electrodes conceived and applied on substrate. Then Gold (Au) is deposited by vacuum deposition to form drain, source and gate electrodes. A 200 nm thick layer of Gold is obtained.
By ink-jetting Orgacon™ IJ-1005 ink (supplied by AGFA), a PEDOT:PSS layer is deposited on source electrode, drain electrode and between electrodes so as to form channel (3). A thermal treatment of 60 minutes at 120° C. is applied to get conductive properties of PEDOT:PSS.
A mixture of 0.2% wt platinum nanoparticles with a 4-(trimethylammonium)thiophenolate crown in PEDOT:PSS is prepared and ink-jetted on a part of gate electrode (2G), yielding a catalytic layer (5) of thickness 3 μm.
Electrodeposition of Biologic Detection Layer
A solution comprising 1.5 wt % Chitosan (75%-85% deacetylated natural chitin, medium molecular weight, MW 190-310 kDa, from Aldrich, CAS 9012-76-4) is prepared in 0.075 M HCl, under vigorous stirring. The resulting solution is filtered over a porosity 3 glass frit to remove any undissolved material. The pH of the solution is adjusted to 5.1 with NaOH 1.0 M. 5.0 mg/mL Glucose Oxidase (Glucose Oxidase type VII from Aspergillus Niger, ≥100 kU/g, MW 160 kDa, from Aldrich) are added to the Chitosan solution under gentle stirring, and 5.0 mL of the resulting mixture are transferred to a 4-way electrochemical cell, equipped with a 3-electrodes setup (working electrode:OECT gate, prepared as previously described; counter electode:inox grid; reference electrode:Saturated Calomel Electrode, Sat'd KCl) and a magnetic stirrer. 20.0 mmol/L H2O2 (H2O2 30 wt % in H2O, from Aldrich) are added to the electrochemical cell. The working electrode is polarized at E=−0.40 V vs SCE, for 200 seconds, at room temperature and under stirring (400 rpm). The working electrode (OECT gate) is then rinsed with distilled water to remove any loosely bound material and stored in air until further use.
OECT-1 devices are thus obtained, as shown schematically on
Preparation of OECT-C1
A comparative OECT device is prepared with the same protocol as OECT-1, but
Only PEDOT:PSS is ink-jetted on a part of gate electrode (2G), yielding a layer (5) of thickness 3 μm. The same amount of PEDOT:PSS is deposited in OECT-C1 and in OECT-1.
Preparation of OECT-C2
A comparative OECT device is prepared with the same protocol as OECT-1 (including electrodeposition of biologic detection layer), but a 0.2% wt platinum nanoparticles with a 4-(trimethylammonium)thiophenolate crown in ethanol is used to prepare catalytic layer (5) on gate electrode (2G). The same amount of nanoparticle solution is deposited in OECT-C2 and in OECT-1.
Preparation of OECT-2
On an OECT-1 device, an encapsulation membrane is further deposited.
A solution comprising 64.0 mg/mL poly-(4-vinylpyridine) (average Mw 160 kDa, from Aldrich) is prepared in a mixture of absolute ethanol and distilled water (90/10 v/v) (solution A). A solution comprising 64.0 mg/mL poly-(sodium 4-styrenesulfonate) (average Mw 1000 kDa, from Aldrich) is prepared in distilled water (solution B). A solution comprising 4.0 mg/mL poly(ethylene glycol) diglycidyl ether (PEGDE, average Mw 500 g/mol, from Aldrich) is prepared in a mixture of absolute ethanol and distilled water (80/20 v/v) (solution C). Solutions A and B are combined with a 90/10 v/v ratio (solution D). Solutions D and C are combined with an 80/20 v/v ratio (solution E). 35.5 μL/cm2 of solution E are drop cast onto the channel and gate of the OECT. The resulting drop is left to dry at room temperature, and under an ethanol/water saturated atmosphere, for 18 to 24 h.
OECT-2 device is thus obtained.
Mode of Operation of OECT—Performances.
OECTs are connected to usual electric device for control of filed effect transistor.
VGS and VDS are set to +0.5V and −0.6V respectively.
A sample containing controlled concentration of glucose in Phosphate Buffer Solution (pH=7.4) is brought in contact with OECT-1. IDS and jGS are measured. Then sample is replaced with a sample more concentrated in glucose.
Setup of example 1 is reproduced, but using OECT-2 device and with following conditions.
A sample of unknown concentration of glucose is used.
OECT is configured in conditions of example 1, then sample is brought in contact with OECT. After few seconds, IDS and jGS becomes stationary. Using sensibility of OECT defined in example 1, real concentration of glucose in sample is determined.
A comparative experiment has been conducted to demonstrate the efficiency of the composite material used in catalytic layer in this disclosure.
OECT-C1 has been used on one hand. In this OECT, there is no biologic detection layer. As a consequence, OECT-C1 is not designed to measure a concentration of glucose, but only a concentration of hydrogen peroxide H2O2.
On the other hand, OECT-1 has been prepared, but without biologic detection layer.
OECT-C1 and OECT-1 have been contacted with H2O2 solutions having a concentration varying from 0.1 mmol/L to 2 mmol/L.
It is clear that the composite material comprising platinum nanoparticles and a PEDOT:PSS conductive matrix has improved sensitivity has compared to PEDOT:PSS conductive matrix alone.
| Number | Date | Country | Kind |
|---|---|---|---|
| 19306167.8 | Sep 2019 | EP | regional |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/EP2020/076605 | 9/23/2020 | WO |
