WO2016176692A2 - Puce de biocapteur - Google Patents

Puce de biocapteur Download PDF

Info

Publication number
WO2016176692A2
WO2016176692A2 PCT/US2016/038186 US2016038186W WO2016176692A2 WO 2016176692 A2 WO2016176692 A2 WO 2016176692A2 US 2016038186 W US2016038186 W US 2016038186W WO 2016176692 A2 WO2016176692 A2 WO 2016176692A2
Authority
WO
WIPO (PCT)
Prior art keywords
biosensor chip
electrode
electrodes
working electrode
thru
Prior art date
Application number
PCT/US2016/038186
Other languages
English (en)
Other versions
WO2016176692A3 (fr
WO2016176692A8 (fr
Inventor
Michael J. Ahrens
Adam G. GAUSTAD
Dimitra Georganopoulou
Yilja Paul BAO
Fang Lai
Rebecca HOO
Original Assignee
Ohmx Corporation
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Ohmx Corporation filed Critical Ohmx Corporation
Priority to PCT/US2016/038186 priority Critical patent/WO2016176692A2/fr
Publication of WO2016176692A2 publication Critical patent/WO2016176692A2/fr
Publication of WO2016176692A8 publication Critical patent/WO2016176692A8/fr
Publication of WO2016176692A3 publication Critical patent/WO2016176692A3/fr

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/403—Cells and electrode assemblies

Definitions

  • aspects described herein relate generally to biosensor chips, and more specifically to biosensor chips that provides conductivity on both sides of the chip, and methods for using biosensor chips to detect electrochemical signals.
  • Biosensor chips are known to have a counter electrode and a working electrode.
  • Conventional standards for biosensor chips recommend that the counter electrode be significantly larger than the working electrode, preferably twice the size of the working electrode, and positioned away from the working electrode.
  • a biosensor chip in an illustrative embodiment, includes a reference electrode, a working electrode having an exposed surface area, and a counter electrode having an exposed surface area.
  • the exposed surface area of the working electrode is larger than the exposed surface area of the counter electrode.
  • a biosensor chip in another illustrative embodiment, includes a reference electrode, a working electrode, and a counter electrode.
  • the working electrode is spaced from the counter electrode by a gap of between and including 0.025 to 0.075 mm.
  • a biosensor chip in yet another illustrative embodiment, includes a reference electrode, a working electrode, and a counter electrode.
  • the reference, working and counter electrodes are shaped and positioned to combine to form a substantially circular shape.
  • a biosensor chip in yet another illustrative embodiment, includes a base material having a first protruding tab, a second protruding tab, and a third protruding tab.
  • the biosensor chip also includes a reference electrode, a working electrode, and a counter electrode positioned on the base material.
  • a biosensor chip includes a base material having a first protruding tab and a second protruding tab, the first and second tabs being different sizes.
  • the biosensor chip also includes a reference electrode, a working electrode and a counter electrode positioned on the base material.
  • a biosensor in another illustrative embodiment, includes a base material and three electrodes disposed on the base material.
  • the biosensor chip also includes thru- holes containing (e.g. filled or internally coated with) a conductive material, with each electrode having at least one thru -hole, and the thru -holes passing through the base material.
  • the biosensor chip also has conductive pads surrounding the thru-holes, with each electrode having at least one conductive pad.
  • the method includes attaching the biosensor chip to a holey container to form a detection well, adding an electrically conductive solution to the detection well such that all three electrodes are immersed in the electrically conductive solution, connecting the detection well with a potentiostat through interconnected conductive alignment pads and thru-hole pads each corresponding to an electrode and detecting signals from the interconnected conductive alignment pads to determine proper electrical connection to the
  • the method includes attaching a biosensor chip having three electrodes to a holey container to form a detection well, adding an assay mix containing electroactive molecules and an electrically conductive solution to the detection well such that the three electrodes are immersed in the electrically conductive solution and a self-assembled monolayer is formed by the electroactive molecules, connecting the detection well with a potentiostat through interconnected conductive alignment pads and thru-hole pads each corresponding to an electrode, detecting signals from the interconnected alignment pads to determine proper electrical connection to the potentiostat and detecting signals from the thru-hole pads to measure electrochemical signals from the self-assembled monolayer of the electroactive molecule.
  • FIG. 1 is a schematic of one embodiment of a front side of a biosensor chip
  • FIG. 2 is a schematic of a back side of the biosensor chip of FIG. 1;
  • FIG. 3 is a schematic of the features of the back side of the biosensor chip of FIG. 1 superimposed onto the features of the front side of the biosensor chip to provide a composite view of all features of both sides of the biosensor chip;
  • FIG. 4 is the composite view of FIG. 3 with certain dimensions annotated
  • FIG. 5 also is the composite view of FIG. 3 with certain dimensions annotated
  • FIG. 6 is an isometric view of a holey container that can be used with a biosensor chip according to one aspect
  • FIG. 7 is a bottom view of the holey container of FIG. 6;
  • FIG. 8 is a cyclic voltammetry profile of a double-sided biosensor chip immersed in an electrolyte solution
  • FIG. 9 depicts cyclic voltammetry profiles of a double-sided biosensor chip immobilized with self-assembled monolayers of hydrogen peroxide sensitive electroactive moiety with hydrogen peroxide (solid lines) or without hydrogen peroxide (diamond lines);
  • FIG. 10 depicts peak ratios of the biosensor chip with self-assembled monolayers exposed to different amounts of hydrogen peroxide.
  • Electron transfer reactions are critical steps in a wide variety of biological interactions ranging from photosynthesis to aerobic respiration. Studies of electron transfer reactions in both chemical and biological systems have yielded a strong theoretical base which describes the rate of electron transfer in terms of a small number of parameters. With this knowledge, an electroactive moiety (EAM) may be designed as a sensing molecule. The magnitude of electron transfer is changed upon its interaction with a target.
  • EAM electroactive moiety
  • Electron transfer is generally initiated electronically, with voltage application being preferred. Precise control and variations in the applied potential can be achieved via a potentiostat and with either a three electrode system (one reference electrode, one working electrode, and one counter electrode) or a two electrode system (one working electrode and one counter electrode).
  • the "counter electrode” is sometimes referred to as an "auxiliary electrode.” This arrangement allows matching of applied potential to peak electron transfer potential of the system, which depends in part on the choice of redox active molecules and in part on the conductive oligomer used.
  • the working electrode is typically made of an "inert" material such as gold.
  • the WE serves as a surface on which the electrochemical reaction takes place.
  • the WE also may serve as an attachment surface for monolayers or self-assembled monolayers (SAM) when such monolayers are involved in the electrochemical reaction and/or
  • the reference electrode (RE) is used as an internal or in-system calibration point to measure the working electrode potential.
  • the RE should have a constant electrochemical potential as long as no current flows through it, allowing the WE potential to be measured/calculated against it.
  • Silver-silver chloride (Ag/AgCl) is a common choice for reference electrode material.
  • the counter electrode (CE) serves as a source or sink for electrons so that current can be passed from an external circuit through an assay well or a detection well - a collection of electrodes and container which can hold an electrolyte or electrically conductive solution.
  • Conventional standards recommend that the counter electrode be significantly larger than the working electrode - preferably twice the size of the working electrode - and positioned away from the working electrode (Ramsay, Graham. Commercial
  • Biosensors Applications to Clinical, Bioprocess, and Environmenta Samples, Wiley, March 16, 1998, Science, 304 pages.
  • the inventors have recognized that, due to these restrictions, the size of the working electrode is limited to a defined space of the whole electrode system, particularly if the electrode system is to be confined to an assay well. This size restriction can affect the signal strength in an assay when a large working electrode is desired.
  • biosensor chips including both two and three electrode systems, are onesided with all electrodes on the "front" side of a base material.
  • Base materials are typically made of an insulating polymer. All of the electrodes are immersed in an electrolyte contained in a container, hereinafter referred to as an assay well or detection well. In this configuration, electrodes or leads connected to electrodes need to be extended out from the assay well to connect to a potentiostat from the front side, making it difficult to have a multi-well format.
  • Thru-hole technology has also been used to make biosensor chips.
  • Thru-hole technology was initially used for electronic components that involve the use of leads on the components that are inserted into holes drilled in printed circuit boards (PCB) and soldered to pads on the opposite side.
  • PCB printed circuit boards
  • pads or conductive pads which surround the hole on both sides.
  • a connection to the electrode is established through these pads.
  • the ability to connect to a potentiostat from the opposite side of the main electrode through pads connected via thru-holes is especially useful for a multi-well format.
  • PCB printed circuit boards
  • the biosensor chip has conductivity on both sides, and therefore is referred to as a double-sided biosensor chip. Methods of using the biosensor chip in a bioelectronic assay for detecting a target analyte in a sample is also described herein.
  • Electrodes refers to a composition which, when connected to an electronic device, is able to sense a current or charge and convert it to a signal. Electrodes may include, but are not limited to, certain metals and their oxides, including gold, platinum, palladium, silicon, aluminum; metal oxide electrodes including platinum oxide, titanium oxide, tin oxide, indium tin oxide, palladium oxide, silicon oxide, aluminum oxide, molybdenum oxide (Mo 2 0 6 ), tungsten oxide (W0 3 ) and ruthenium oxides; and carbon, including glassy carbon electrodes, graphite, and carbon paste. Electrodes also may include gold, silicon, carbon, and metal oxide electrodes, with gold being preferred in some embodiments.
  • the working electrode of the biosensor chip is the largest electrode in a three electrode system.
  • three electrodes are arranged with the working electrode being the largest electrode, and being in the center and flanked by the counter electrode on one side and the reference electrode on the other side.
  • the reference electrode is smaller than the counter electrode.
  • the reference electrode is made of or covered with silver-silver chloride (Ag/AgCl).
  • the three electrodes may be shaped so as to maximize the surface area available within a given spatial limitation, for example, the spatial limitation of an assay well.
  • each electrode may vary depending on applications. For example, flat planar electrodes may be preferred for additional optical detection when, for example, arrays of nucleic acids are made, thus requiring addressable locations for both synthesis and detection.
  • the electrode may be in the form of a tube to increase surface area.
  • electrodes are formed on a base material.
  • the base material may be an insulating substrate.
  • the insulating substrate may be, but is not limited to, a polymer. Different processes such as lithography or laser ablation may be used to form the patterns of electrodes, the leads that connect the electrodes to other conductive features, as well as conductive features such as extended areas.
  • a thru-hole is made through individual electrodes by making a hole through the electrode and the base material.
  • the hole is sub-millimeters in diameter.
  • a conductive material such as conductive silver or carbon is deposited onto the base material of the biosensor chip to fill the hole.
  • a conductive material such as silver or carbon is deposited onto the base material of the biosensor chip to create a pad surrounding the hole. In some embodiments, both are done simultaneously with the same material. In this manner, an electrode on one side of the biosensor chip is connected to a pad on the other side of the biosensor chip via the thru-hole containing the conductive material.
  • connection to a potentiostat is easily achieved by connecting the potentiostat to one or more pads on the back side of the chip. Suitable printing methods for depositing conductive materials known to those skilled in the art may be used.
  • additional conductive pads and leads which are not associated with a thru-hole are printed on the back side of the biosensor chip for a connection check. These pads are hereinafter referred to as alignment pads. Proper alignment of the alignment pads relative to connecting leads, wires, and/or probes of a potentiostat, forms an electrical circuit, thereby establishing the connection between the biosensor chip and the potentiostat.
  • the alignment pads maybe used to determine whether a proper connection has been made to the potentiostat.
  • the biosensor chip can be in a variety of shapes, e.g., round, square, irregular, symmetric or asymmetric. In some embodiments, the biosensor chip may include protruding tabs that extend off of the main shape.
  • the shape of the biosensor chip is generally round with three extended tabs of different sizes.
  • Each tab may serve as a space for thru-holes and conductive pads.
  • the electrodes or leads from the electrodes may extend onto the tabs.
  • biosensor chips may be incorporated into a cartridge and may take a variety of configurations.
  • the chips may be part of reaction chambers with inlet and outlet ports for the introduction and removal of reagents.
  • the cartridges may include caps or lids that have microfluidic components, so that the sample can be introduced, reagents can be added, reactions can occur, and then the reaction mix can be added to the reaction chamber for detection.
  • the biosensor chip is adhered to the opened end of a hollow tube or holey container such that the biosensor chip forms a solidly attached end-cap on one end of the tube or holey container in such a way that the electrodes can be immersed in an electrolyte or other electrically conductive solution.
  • the combination of the biosensor chip with the container e.g. a holey container
  • the container e.g. a holey container
  • an electrically conductive solution can be referred to as a detection well.
  • the holey container has an inner dimension that is slightly larger than the outer dimensions of the three electrodes so that the electrodes can be completely immersed in an electrically conductive solution.
  • the wall of the holey container is thick enough to provide sufficient surface area for a firm attachment of the holey container to the biosensor chip.
  • the bottom shape of the holey container mirrors the shape of the biosensor chip so that the holey container and the chip only have a single alignment orientation. Assembly processes such as laser welding and use of pressure sensitive adhesive followed by UV cure, may be used to attach a biosensor chip to a container to form an assay well or detection well.
  • a biosensor chip may have one or more and any combination of the features described above. Contrary to conventional designs, in some
  • the biosensor chip has a working electrode with surface area larger than the counter electrode, with the working and counter electrodes spatially close together, yet is still functional.
  • the biosensor chip includes a reference electrode without a significant reduction of the area of the working electrode. Such an arrangement may make signal outputs higher, making measurements easier in some cases.
  • the biosensor chip has at least one thru-hole per electrode containing conductive materials, allowing for electrical connection and detection from the back side, specifically the bottom side of the electrodes, thereby simplifying the design of the detection well and the electronic device.
  • the detection chip does not necessarily have extended (often thin and flexible) electrodes protruding from the detection well for connection purposes. With the thru-holes, the assembled detection well can simply and firmly "sit" on a post or probe for connection to a potentiostat.
  • the biosensor chip has an asymmetric shape for proper orientation during assembly of the detection well.
  • the biosensor chip has two interconnected alignment pads on the back side of the chip for detection of proper connectivity to a potentiostat.
  • the biosensor chip described herein can be used for sensing electrochemical signals in many ways. Once assembled with a holey container or other container to form a detection well, the chip may be used to directly measure the conductivity of an electrically conductive solution. In some cases, the chip may be used indirectly to detect a target analyte in a sample through the formation of a self-assembled monolayer (SAM) comprising an electroactive moiety (EAM) on the electrodes before or after the EAM is exposed to a reaction mix containing the target analyte or a surrogate target analyte such that the target or surrogate target interacts with the EAM and a signal proportional to the target concentration is obtained.
  • SAM self-assembled monolayer
  • EAM electroactive moiety
  • FIG. 1 depicts a front side of a biosensor chip 1 according to one illustrative embodiment.
  • the biosensor chip 1 comprises a base material 10 and three electrodes disposed on the base material: a counter electrode 21, a reference electrode 22 and a working electrode 23.
  • the counter electrode 21 is shaped as a circular segment
  • the reference electrode 22 is shaped as a circular segment
  • the working electrode 23 is shaped as a portion of a circle that lies between two parallel lines cutting the circle. It should be understood that the electrodes may be any other suitable shape.
  • Each of the electrodes has a corresponding lead that connects the electrode out to a conductive extended area.
  • a first lead 31 connects the counter electrode 21 out to a corresponding first extended area 41
  • a second lead 32 connects the reference electrode 22 to a corresponding second extended area 42
  • a third lead 33 connects the working electrode 23 out to a corresponding third extended area 43.
  • the electrode leads each have a thickness of between and including 0.25 to 1.25 mm, or 0.5 to 1 mm. In some embodiments, the electrode leads may have a thickness of 0.75 mm.
  • the extended areas may be made of any suitable conductive material.
  • Each of the extended areas has one or more thru-holes 30 that extend through the base material 10 from the front side of the biosensor chip to a back side of the biosensor chip.
  • Each extended area may have one, two, three, or any suitable number of thru-holes.
  • the back side of the biosensor chip is depicted in FIG. 2.
  • the back side of the biosensor has conductive pads 81, 82 and 83.
  • each conductive pad on the back side of the biosensor chip corresponds with an extended area on the front side of the chip and may be aligned with that corresponding extended area.
  • a first conductive pad 81 on the back side of the biosensor chip is aligned with its corresponding first extended area 41 on the front side of the biosensor chip.
  • a second conductive pad 82 is aligned with its corresponding second extended area 42
  • a third conductive pad 83 is aligned with its corresponding third extended area 43.
  • the thru-holes 30 extend from an extended area to a corresponding conductive pad.
  • the thru-holes contain a conductive material to allow an electrical connection to be made between the electrodes on the front side of the biosensor chip and the conductive pads on the back side of the biosensor chip.
  • Each conductive pad is aligned with its corresponding extended area such that the two areas overlap.
  • FIG. 3 depicts the features of the back side of the biosensor chip of FIG. 1 superimposed onto the features of the front side of the biosensor chip to provide a composite view of all features of both sides of the biosensor chip.
  • each pair of conductive pad and corresponding extended area overlap one another such that thru- holes can pass through both of them.
  • first extended area 41 overlaps with first conductive pad 81
  • second extended area 42 overlaps with second conductive pad 82
  • third extended area 43 overlaps with third conductive pad 83.
  • the conductive pads and extended areas may also be referred to herein as "thru-hole pads.”
  • the conductive pads and extended areas may be circular, square, rectangular, oval-shaped, or any other suitable shape.
  • the base material of the biosensor chip has one or more protruding tabs.
  • the base material 10 has three protruding tabs 11, 12, 13.
  • the tabs may each comprise an arc shape.
  • the tabs may have a different shape such as a rectangle.
  • the protruding tabs are each a different size than the other protruding tabs.
  • each of the protruding tabs has an arc length different from the other two, making the shape of the base material 10 asymmetric. As discussed above, this asymmetric shape may be used to help properly align the biosensor chip with a corresponding holey container.
  • the back side of the biosensor chip also includes alignment pads 61, 62, which are connected to one another via a lead 70.
  • the electrodes of the biosensor chip are connected to a potentiostat via the leads and conductive extended areas on the front side of the chip, the conductive material located in the thru -holes, and the conductive pads located on the back side of the chip. Detection of signals from the conductive extended areas is used to measure electrochemical signals from the solution.
  • the interconnected alignment pads are also connected to the potentiostat. Detection of signals from the interconnected alignment pads determines proper electrical connection between the biosensor chip and the potentiostat.
  • Each of the protruding tabs may have an extended area and corresponding conductive pad disposed on the tab.
  • first extended area 41 is disposed on first tab 11
  • second extended area 42 is disposed on second tab 12
  • third extended area 43 is disposed on third tab 13.
  • each of the alignment pads is disposed upon a protruding tab.
  • An alignment pad may share a protruding tab with a conductive pad.
  • the first alignment pad 61 is disposed on the first tab 11, and shares the first tab 11 with the first conductive pad 81.
  • the second alignment pad 62 is disposed on the second tab 12, and shares the second tab 12 with the second conductive pad 82.
  • FIGS. 4-5 depict a composite view of the biosensor in which features on the back side of the biosensor chip superimposed onto the features of the front side of the biosensor chip, with certain dimensions annotated.
  • the working and counter electrodes of the biosensor chip are spatially close together, contrary to conventional standards. As seen in FIG. 4, there is a gap 51 between the counter electrode 21 and the working electrode 23. In some embodiments, the gap between the counter electrode and the working electrode is between and includes 0.01 to 0.09 mm, 0.025 to 0.075 mm, 0.04 to 0.06 mm, 0.045 to 0.055 mm, or is 0.05 mm.
  • a gap 52 also exists between the reference electrode 22 and the working electrode 23.
  • the gap between the reference electrode and the working electrode is between and includes 0.25 to 1.25 mm, 0.5 mm to 1 mm, 0.65 to 0.85 mm, 0.7 to 0.8 mm, or is 0.75 mm.
  • FIG. 4 depicts heights of counter electrode 21 and the reference electrode 22.
  • the height HI of the counter electrode 21 is between and includes 1 to 2 mm, 1.25 to 1.75 mm, or is 1.5 mm.
  • the height H2 of the reference electrode 22 is between and includes 0.6 to 1.6 mm, 0.8 to 1.4 mm, or is 1.1 mm.
  • FIG. 5 depicts the relative angles between each of the extended
  • the angles are measured from the center points of each of these features.
  • the angle ⁇ 2 between the second extended area 42 and the third extended area 43 is between and includes 90 to 120 degrees, 95 to 115 degrees or is 105 degrees.
  • the angle between the first extended area 41 and the second extended area 42 is between and includes 140 to 160 degrees, 145 or 155 degrees, or is 150 degrees.
  • the angle between the first extended area 41 and the third extended area 43 is between and includes 90 to 120 degrees, 95 to 115 degrees or is 105 degrees. It should be understood that the sum of these three angles do not exceed 360 degrees. In some embodiments, these three angles are chosen such that the sum of these three angles is 360 degrees.
  • the angle from the first extended area 41 to the third extended area 43 is the same as the angle from the third extended area 43 to the second extended area, and, in some embodiments, these angles may each be smaller than the angle from the first extended area to the second extended area.
  • the angle ⁇ between the alignment pads 61, 62 is between and includes 80 to 100 degrees, 85 to 95 degrees, or is 90 degrees. In some embodiments, the angle ⁇ between the first extended area 41 to the first alignment pad 61is between and includes 20 to 40 degrees, 25 to 35 degrees, or is 30 degrees. In some embodiments, the angle ⁇ 2 between the second extended area 42 to the second alignment pad 62 is between and includes 20 to 40 degrees, 25 to 35 degrees, or is 30 degrees. In some embodiments, these two angles ⁇ , ⁇ 2 are the same. In some embodiments, the sum of angles ⁇ , ⁇ 2 and ⁇ is between and including 140 to 160 degrees. In some embodiments, angles ⁇ , ⁇ 2 and ⁇ add up to 150 degrees.
  • the protruding tabs 11, 12 and 13 each has an arc length different from the other two, making the shape of the base material 10 asymmetric.
  • the degree measure ⁇ 1 of the arc of the first tab 11 is between and includes 46 to 66 degrees, 50 to 62 degrees, or is 56 degrees.
  • the degree measure ⁇ 2 of the arc of the second tab 12 is between and includes 56 to 76 degrees, 60 to 72 degrees, or is 66 degrees.
  • the degree measure ⁇ 3 of the arc of the third tab 13 is between and includes 36 to 56 degrees, 40 to 52 degrees, or is 46 degrees.
  • the degree measure of the arc of each tab increases by 10 degrees as one moves from the third tab 13 to the first tab 11 to the second tab 12.
  • the base material 10 also includes arc segments extending between each consecutive pair of protruding tabs.
  • a first arc segment 121 extends between the first tab 11 and the second tab 12
  • a second arc segment 122 extends between the second tab 12 and the third tab 13
  • a third arc segment 123 extends between the third tab 13 and the first tab 11.
  • the degree measure al of the first arc segment 121 is between and includes 49 to 69 degrees, 55 to 65 degrees, or is 59 degrees.
  • the degree measure a2 of the second arc segment 122 is between and includes 54 to 74 degrees, 59 to 69 degrees, or is 64 degrees.
  • the degree measure a3 of the third arc segment 123 is between and includes 59 to 79 degrees, 65 to 75 degrees, or is 69 degrees. In some embodiments, the degree measure of each arc segment increases by 5 degrees as one moves from the first arc segment 121 to the second arc segment 122 to the third arc segment 123. In some embodiments, sum of all arc segments of the protruding tabs and all arc segments extending between each consecutive pairs of protruding tabs (i.e., ⁇ 1+ ⁇ 2+ ⁇ 3+ ⁇ 1+ ⁇ 2+ ⁇ 3) totals 360 degrees.
  • the distance from the center of the base material 10 to the arc segments between the protruding tabs is represented by Rl.
  • Rl is between and includes 3 to 5 mm, 3.7 to 4.7 mm, or is 4.2 mm.
  • the distance from the center of the base material 10 to the centers of each of the extended areas, conductive pads, and alignment pads is the same. In FIG. 4, this distance is represented by R2. In some embodiments, R2 is between and includes 4.2 to 6.2 mm, 4.5 to 5.9 mm, or is 5.2 mm.
  • the distance from the center of the base material 10 to the radially outward edge of each of the protruding tabs is the same. In FIG. 4, this distance is represented by R3. In some embodiments, R3 is between and includes 5.2 to 7.2 mm, 5.7 to 6.7 mm, or is 6.2 mm.
  • an angle ⁇ 1 exists between the third extended area 43 and the side edge of the third tab 13.
  • ⁇ 1 is between and includes 13 to 33 degrees, 18 to 28 degrees or is 23 degrees.
  • a biosensor chip is attached to a holey container to form a detection well.
  • the holey container may be placed on top of the front side of the biosensor chip to form a vessel into which an electrically conductive solution may be added.
  • FIGS. 6 and 7 depicts a bottom view.
  • the holey container 100 includes an opening 110 that receives electrically conductive solution and an outer wall 130.
  • Protruding tabs 111, 112 and 113 protrude from the bottom of the outer wall 130.
  • the size, shape and position of the protruding tabs of the holey container maybe designed to match the size, shape and position of the protruding tabs of a corresponding biosensor chip.
  • the holey container 100 shown in FIGS. 6-7 is designed to correspond to the shape of the biosensor chip of FIG. 1.
  • first tab 111, second tab 112, and third tab 113 of the holey container 100 are sized, shaped and positioned to align with the first tab 11, second tab 12, and third tab 13 of the biosensor chip 1 when the holey container 100 is placed on top of the front side of the biosensor chip.
  • an electrically conductive solution may be added to the detection well to immerse the electrodes of the biosensor chip in the solution.
  • the detection well is connected to a potentiostat via the interconnected conductive alignment pads and the thru-holes running through the extended area/conductive pad pairs, where each pair corresponds to an electrode.
  • the dashed lines in FIG. 4 represent the boundaries of the holey container with the holey container placed on top of the front side of the biosensor chip, the inner wall of the holey container is spaced from the electrodes of the biosensor chip.
  • the innermost dashed line represents the location of the inner wall 131 of the holey container when the holey container is placed on top of and properly aligned with the biosensor chip.
  • a gap 53 exists between the outer circumference of the electrodes and the inner wall of the holey container. In some embodiments, this gap 53 is between and includes 0.3 to 0.5 mm, 0.35 to 0.45 mm, or is 0.39 mm. As also seen in FIG. 4, there is a distance 54 from the inner wall 131 of the holey container to the arc segments 121, 122, 123 of the base material 10. In some embodiments, this distance 54 is between and includes 0.8 to 1.2 mm, 0.9 to 1.1 mm, or is 1 mm.
  • the chip is in general based on a circular shape with three tabs having differing arc lengths (FIG. 1).
  • the base material of the chip is made of polystyrene.
  • the electrodes, leads and extended areas (FIG. 1) are made of gold.
  • the reference electrode is further coated with silver-silver chloride. Thru-holes are created, two for each electrode, in the extended areas and contain conductive silver (FIG.l).
  • On the back side of the chip (FIG. 2), five pads are printed with conductive silver.
  • conductive pads Three of the pads, called conductive pads, overlap with the extended areas of electrodes, allowing the conductive silver to flow through the thru-holes and cover at least a portion of the extended areas on the front side such that an electrical connection can be made between the electrodes on the front side and the pads on the back side.
  • Two of the pads, called alignment pads are interconnected with one another and aid in alignment control. These alignment pads verify proper connection with a potentiostat.
  • a detection well is formed by attaching the holey container to the front side of the biosensor chip. An electrically conductive solution is added to the detection well, allowing the three electrodes of the biosensor chip to be immersed the solution.
  • the assembled detection well is connected to a potentiostat through the pads on the back side of the biosensor chip. Proper connection to the potentiostat is monitored through the two alignment pads.
  • FIG. 8 depicts a cyclic voltammetry profile of the biosensor chip.
  • the solid line outlines the resulting cyclic voltammetry scanning profile from the chip.
  • the profiles demonstrate a single peak (in solid lines) from the biosensor chips with the SAM not being exposed to H 2 0 2 and double peaks (in diamond lines) from the biosensor chips with the SAM being exposed to lOOuM H 2 0 2 , indicating that the biosensor chips are fully functional and differential responses can be measured from the back side of the biosensor chip with its thru-hole design.
  • FIG. 10 depicts peak ratios of the electrode chips with SAM exposed to 0 ⁇ H 2 O 2 or to 100 ⁇ H 2 O 2 .
  • Peak current ratios (Ip 2 / Ipi) show a clear difference between 0 ⁇ and 100 ⁇ H 2 0 2 , where I P i is the total current of the top peak observed at the more positive voltage potential and I P2 is the total current of the top peak observed at the more negative voltage potential.
  • I P i is the total current of the top peak observed at the more positive voltage potential
  • I P2 is the total current of the top peak observed at the more negative voltage potential.
  • Each standard deviation bar shown is calculated from quadruplicate.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Electrochemistry (AREA)
  • Molecular Biology (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Investigating Or Analyzing Materials By The Use Of Electric Means (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

Cette invention concerne une puce de biocapteur destinée à détecter des signaux électrochimiques. Selon certains modes de réalisation, ladite puce de biocapteur est biface et elle comprend trois électrodes. Selon certains modes de réalisation, ladite puce de biocapteur comprend une électrode de travail et une contre-électrode qui sont spatialement rapprochées. Selon certains modes de réalisation, le matériau de base de la puce de biocapteur présente trois languettes saillantes dont chacune présente une taille différente par rapport aux autres languettes saillantes. Ledit biocapteur peut être combiné avec un récipient perforé pour former un puits de détection.
PCT/US2016/038186 2016-06-17 2016-06-17 Puce de biocapteur WO2016176692A2 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
PCT/US2016/038186 WO2016176692A2 (fr) 2016-06-17 2016-06-17 Puce de biocapteur

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/US2016/038186 WO2016176692A2 (fr) 2016-06-17 2016-06-17 Puce de biocapteur

Publications (3)

Publication Number Publication Date
WO2016176692A2 true WO2016176692A2 (fr) 2016-11-03
WO2016176692A8 WO2016176692A8 (fr) 2016-12-08
WO2016176692A3 WO2016176692A3 (fr) 2017-01-05

Family

ID=57199417

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2016/038186 WO2016176692A2 (fr) 2016-06-17 2016-06-17 Puce de biocapteur

Country Status (1)

Country Link
WO (1) WO2016176692A2 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111007129A (zh) * 2019-12-17 2020-04-14 深圳市刷新智能电子有限公司 一种石墨烯生物传感器电极的制备工艺

Family Cites Families (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2795315B2 (ja) * 1996-05-16 1998-09-10 日本電気株式会社 半導体装置
US6134461A (en) * 1998-03-04 2000-10-17 E. Heller & Company Electrochemical analyte
DE69931880T2 (de) * 1998-12-14 2007-05-31 Eastman Kodak Co. Elektrische Verdrahtung hoher Dichte für einen kontinuierlich arbeitenden Tintenstrahldruckkopf
AU2001261145B2 (en) * 2000-05-03 2005-08-11 The United States Government, As Represented By The Department Of The Navy Biological identification system with integrated sensor chip
US6908470B2 (en) * 2001-01-11 2005-06-21 Fraunhofer-Gesellschaft Zur Foderung Der Angewandten Forschung E.V. Sieve electrode which can be connected to a nerve stump
US6756296B2 (en) * 2001-12-11 2004-06-29 California Institute Of Technology Method for lithographic processing on molecular monolayer and multilayer thin films
CA2690190A1 (fr) * 2007-07-02 2009-01-08 Genefluidics, Inc. Dosage sur puce ayant une efficacite amelioree
US20100227080A1 (en) * 2007-10-05 2010-09-09 Beer Greg P Method of Defining Electrodes Using Laser-Ablation and Dielectric Material
US8552548B1 (en) * 2011-11-29 2013-10-08 Amkor Technology, Inc. Conductive pad on protruding through electrode semiconductor device

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111007129A (zh) * 2019-12-17 2020-04-14 深圳市刷新智能电子有限公司 一种石墨烯生物传感器电极的制备工艺
CN111007129B (zh) * 2019-12-17 2022-05-13 深圳市刷新智能电子有限公司 一种石墨烯生物传感器电极的制备工艺

Also Published As

Publication number Publication date
WO2016176692A3 (fr) 2017-01-05
WO2016176692A8 (fr) 2016-12-08

Similar Documents

Publication Publication Date Title
Rahn et al. Recent advancements in bipolar electrochemical methods of analysis
Arora et al. A wireless electrochemiluminescence detector applied to direct and indirect detection for electrophoresis on a microfabricated glass device
Ozkan et al. From mercury to nanosensors: Past, present and the future perspective of electrochemistry in pharmaceutical and biomedical analysis
Privett et al. Electrochemical sensors
Uslu et al. Electroanalytical methods for the determination of pharmaceuticals: a review of recent trends and developments
Wei et al. Electrochemical biosensors at the nanoscale
JP3672525B2 (ja) バイオセンサー
RU2490622C2 (ru) Тестовые датчики, способы и системы с множеством зон и потенциалов
CN1205474C (zh) 生物传感器
EP3926339B1 (fr) Capteur électrochimique comportant des zones sensibles aux analytes distribuées symétriquement
JP2005530179A (ja) 多成分の微量分析のための微細加工で作られたセンサーアレイ
US8202408B2 (en) Sensor electrode and method for the electrochemical detection of nucleotides
EP2005156A2 (fr) Biocapteur miniaturise avec detection amperometrique optimisee
WO2018014719A1 (fr) Structure d'électrode de détection, plaque d'ouverture de détection, et plaque d'ouverture de détection préfabriquée
US9671432B2 (en) Nanowire electrode sensor
WO2005033685A2 (fr) Plates-formes de capteurs utilisant des membranes nanoporeuses
Deroco et al. based electrochemical sensing devices
Gillis et al. Electrochemical measurement of quantal exocytosis using microchips
Muratova et al. Voltammetric vs. potentiometric sensing of dopamine: advantages and disadvantages, novel cell designs, fundamental limitations and promising options
Sripirom et al. Simple and economical analytical voltammetry in 15 μl volumes: Paracetamol voltammetry in blood serum as a working example
Kim et al. A miniaturized electrochemical system with a novel polyelectrolyte reference electrode and its application to thin layer electroanalysis
WO2016176692A2 (fr) Puce de biocapteur
Tadi et al. Voltammetric determination of pindolol in biological fluids using molecularly imprinted polymer based biomimetic sensor
Valentini et al. Fullerene black modified screen printed electrodes for the quantification of acetaminophen and guanine
Choi et al. Electrochemical gene detection using multielectrode array DNA chip

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 16787316

Country of ref document: EP

Kind code of ref document: A2

NENP Non-entry into the national phase in:

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 16787316

Country of ref document: EP

Kind code of ref document: A2