Source: http://www.google.com/patents/US7998325?dq=6,250,774
Timestamp: 2013-12-05 10:15:46
Document Index: 765858518

Matched Legal Cases: ['art.\n13', 'Application No. 2000', 'Application No. 2000', 'Application No. 2000', 'Application No. 2000', 'Application No. 2000', 'art 9', 'art 9', 'art 71', 'art 57', 'art 72', 'art 58', 'art 73', 'art 59', 'art 4117', 'art 4117', 'Application No. 00974977', 'Application No, 09177290']

Patent US7998325 - Biosensor, thin film electrode forming method, quantification apparatus, and ... - Google PatentsSearch Images Maps Play YouTube News Gmail Drive More »Advanced Patent Search | Sign inAdvanced Patent SearchPatentsA biosensor is disclosed comprising a support; a conductive layer composed of an electrical conductive material such as a noble metal, for example gold or palladium, and carbon; slits parallel to and perpendicular to the side of the support; working, counter, and detecting electrodes; a spacer which...http://www.google.com/patents/US7998325?utm_source=gb-gplus-sharePatent US7998325 - Biosensor, thin film electrode forming method, quantification apparatus, and quantification methodPublication numberUS7998325 B2Publication typeGrantApplication numberUS 10/809,217Publication dateAug 16, 2011Filing dateMar 25, 2004Priority dateNov 15, 1999Also published asCN1220053C, CN1340159A, CN1632553A, CN1632554A, CN1632555A, CN100347537C, CN100363739C, EP1152239A1, EP1152239A4, EP2151683A2, EP2151683A3, EP2267439A1, EP2275807A1, US6875327, US8025780, US8142629, US8349157, US8470162, US8475646, US8480866, US8480867, US8480878, US20040178066, US20040178067, US20090152111, US20100243443, US20110117269, US20110147234, US20110174613, US20110272296, US20110278167, US20120251708, WO2001036953A1Publication number10809217, 809217, US 7998325 B2, US 7998325B2, US-B2-7998325, US7998325 B2, US7998325B2InventorsShoji Miyazaki, Hiroyuki Tokunaga, Masaki Fujiwara, Eriko Yamanishi, Yoshinobu TokunoOriginal AssigneePanasonic CorporationPatent Citations (86), Non-Patent Citations (16), Referenced by (7), Classifications (10), Legal Events (1) External Links: USPTO, USPTO Assignment, EspacenetBiosensor, thin film electrode forming method, quantification apparatus, and quantification methodUS 7998325 B2Abstract A biosensor is disclosed comprising a support; a conductive layer composed of an electrical conductive material such as a noble metal, for example gold or palladium, and carbon; slits parallel to and perpendicular to the side of the support; working, counter, and detecting electrodes; a spacer which covers the working, counter, and detecting electrodes on the support; a rectangular cutout in the spacer forming a specimen supply path; an inlet to the specimen supply path; a reagent layer formed by applying a reagent containing an enzyme to the working, counter, and detecting electrodes, which are exposed through the cutout in the spacer; and a cover over the spacer. The biosensor can be formed by a simple method, and provides a uniform reagent layer on the electrodes regardless of the reagent composition.
the electrode part being dividedly formed by a first type of slits provided on an electrical conductive layer which is formed on the whole or part of an internal surface of one or both of the first insulating support and the second insulating support,
a second type of slits is provided around a position where the reagent is dripped on the electrode part so as to partly surround the dripped position and be in contact with the reagent, said second type of slits restricting the spread of the reagent.
2. The biosensor as defined in claim 1, wherein the second type of slits is arc shaped.
3. The biosensor as defined in claim 1, wherein the electrode part further comprises a detecting electrode.
4. The biosensor as defined in claim 3, wherein the counter electrode is provided on the whole or part of the internal surface of the second insulating support,
the working electrode and the detecting electrode which are provided on the internal surface of the first insulating support are dividedly formed by the first type of slits provided on the electrical conductive layer.
5. The biosensor as defined in claim 3, wherein the counter electrode and the detecting electrode have a total area that is equal to or larger than that of the working electrode.
6. The biosensor as defined in claim 5, wherein the area of the detecting electrode in the specimen supply path of the biosensor is equal to the area of the counter electrode.
7. The biosensor as defined in claim 1, wherein the electrode part is provided on the whole or part of the internal surface of only the first insulating support, and the electrode part provided on the internal surface of the first insulating support is dividedly formed by the first type of slits provided on the electrical conductive layer.
8. The biosensor as defined in claim 1, wherein the counter electrode has an area equal to or larger than that of the working electrode.
9. The biosensor as defined in claim 1, wherein a spacer is provided which has a cutout part for forming the specimen supply path and is placed on the electrode part, and the second insulating support is placed on the spacer.
10. The biosensor as defined in claim 9, wherein the spacer and the second insulating support are integral.
11. The biosensor as defined in claim 1, wherein an air hole leading to the specimen supply path is formed.
12. The biosensor as defined in claim 1, wherein a third type of slits is provided for dividing the electrical conductive layer to define an area of the electrode part.
13. The biosensor as defined in claim 12, wherein the first insulating support and the second insulating support are approximately rectangular in shape, and one or more of the third type of slits are provided in parallel to one side of the approximate rectangle shape.
14. The biosensor as defined in claim 1 comprising information of correction data generated for each production lot of the biosensor, which correspond to characteristics concerning output of an electrical change resulting from a reaction between the sample liquid and the reagent layer and can be discriminated by a measuring device employing the biosensor.
15. The biosensor as defined in claim 14, wherein one or a plurality of a fourth type of slits dividing the electrode part are provided, and the measuring device can discriminate the information of the correction data according to positions of the fourth type of slits.
16. The biosensor as defined in claim 1, wherein at least one or all of the first type of slits, the second type of slits which are provided around a position where the reagent is dripped so as to form the reagent layer, the third type of slits which are provided for dividing the electrical conductive layer to define an area of the electrode part, and the fourth type of slits which divide the electrode part, are formed by processing the electrical conductive layer by a laser.
17. The biosensor as defined in claim 16, wherein the first type of slits, the second type of slits, the third type of slits, and the fourth type of slits have a slit width of 0.005 mm to 0.3 mm.
18. The biosensor as defined in claim 1, wherein the reagent layer comprises an enzyme.
19. The biosensor as defined in claim 1, wherein the reagent layer comprises an electron transfer agent.
20. The biosensor as defined in claim 1, wherein the reagent layer comprises a hydrophilic polymer.
21. The biosensor as defined in claim 1, wherein the insulating support is made of a resin material.
This application is a continuation of U.S. patent application Ser. No. 09/889,243, filed Oct. 1, 2001, now U.S. Pat. No. 6,875,327, which is a national stage entry under 35 U.S.C. 371 of PCT International Patent Application No. PCT/JP00/08012, filed Nov. 14, 2000, which claims priority of Japanese patent application Ser. No. 11/324,551, filed Nov. 15, 1999, Japanese Patent Application No. 2000/111255, filed Apr. 12, 2000, Japanese Patent Application No. 2000/113754, filed Apr. 14, 2000, Japanese Patent Application No. 2000/124394, filed Apr. 25, 2000, Japanese Patent Application No. 2000/128249, filed Apr. 27, 2000, and Japanese Patent Application No. 2000/130158, filed Apr. 28, 2000, the contents of all of which are hereby incorporated by reference into the subject application.
TECHNICAL FIELD The present invention relates to a biosensor which quantifies a substrate included in a sample liquid, a thin film electrode forming method suitable at the manufacture of this biosensor, as well as a quantification apparatus and a quantification method using the biosensor and, more particularly, to a biosensor which provides a low manufacture error and a stable performance, a thin film electrode forming method used in manufacturing electrodes of the biosensor, as well as a quantification apparatus and a quantification method using the biosensor.
BACKGROUND ART A biosensor is a sensor which utilizes a molecule recognizing capacity of a biological material such as microorganisms, enzymes, antibodies, DNA, and RNA and applies a biological material as a molecular discrimination element to quantify a substrate included in a sample liquid. That is, the substrate included in the sample liquid is quantified by utilizing a reaction which is caused when a biological material recognizes an objective substrate, such as an oxygen consumption due to respiration of a microorganism, an enzyme reaction, and a luminous reaction. Among various biosensors, an enzyme sensor has progressively come into practical use, and an enzyme sensor as a biosensor for, for example, glucose, lactic acid, cholesterol, and amino acid is utilized in the medical diagnostics or food industry. This enzyme sensor reduces an electron transfer agent by an electron which is generated by a reaction of a substrate included in a sample liquid as a specimen and enzyme or the like, and a quantification apparatus electrochemically measures a reduction quantity of the transfer agent, thereby performing quantitative analysis of the specimen.
The sample liquid (hereinafter, also referred to as �specimen�) is supplied to the inlet 1106 b of the specimen supply path in a state where a fixed voltage is applied between the counter electrode 1103 a and the working electrode 1103 b by a quantification apparatus (hereinafter, also referred to as �measuring device�) connected to the biosensor Z. The specimen is drawn inside the specimen supply path by capillary phenomenon, passes on the counter electrode 1103 a nearer to the inlet 1106 b, and reaches to the working electrode 1103 a, and a dissolution of the reagent layer 1105 is started. At this point of time, the quantification apparatus detects an electrical change occurring between the counter electrode 1103 a and the working electrode 1103 b, and starts a quantification operation. In this way, the substrate included in the sample liquid is quantified.
DISCLOSURE OF THE INVENTION According to the present invention, there is provided a biosensor for quantifying a substrate included in a sample liquid comprising: a first insulating support and a second insulating support; an electrode part comprising at least a working electrode and a counter electrode; a specimen supply path for introducing the sample liquid to the electrode part; and a reagent layer employed for quantifying the substrate included in the sample liquid, and the electrode part, the specimen supply path, and the reagent layer exist between the first insulating support and the second insulating support, the specimen supply path is provided on the electrode part, and the reagent layer is provided on the electrode part in the specimen supply path, respectively, and the electrode part is dividedly formed by first slits provided on an electrical conductive layer which is formed on the whole or part of an internal surface of one or both of the first insulating support and the second insulating support.
According to an embodiment of the present invention, in the biosensor, the electrode part is provided on the whole or part of the internal surface of only the first insulating support, and the electrode part provided on the internal surface of the first insulating support is dividedly formed by the fist slits provided on the electrical conductive layer.
According to an embodiment of the present invention, in the thin film electrode forming method, a degree of the vacuum in the evacuation step is within a range of 1�10−1 to 3�10−3 pascals.
According to an embodimentof the present invention, in the thin film electrode forming method, a degree of the vacuum in the second evacuation step is within a range of 1�10−1 to 3�10−3 pascals.
According to an embodimentof the present invention, in the thin film electrode forming method, the inert gas is either a rare gas of argon, neon, helium, krypton and xenon, or nitrogen.
According to an embodimentof the present invention, in the thin film electrode forming method as defined in any of Claims 29 to 33, the vacuum chamber and the second vacuum chamber is the same chamber.
According to an embodimentof the present invention, in the thin film electrode forming method, the conductive substance is a noble metal or carbon.
According to an embodimentof the present invention, in the thin film electrode forming method, a thickness of a formed thin film electrode is within a range of 3 nm to 100 nm.
According to an embodimentof the present invention, there is provided a quantification method for quantifying, by employing the biosensor, a substrate included in a sample liquid supplied to the biosensor comprising: a fist application step of applying a voltage between the detecting electrode and the counter electrode or the working electrode; a reagent supplying step of supplying the sample liquid to the reagent layer; a first change detecting step of detecting an electrical change occurring between the detecting electrode and the counter electrode or the working electrode by the supply of the sample liquid to the reagent layer; a second application step of applying a voltage between the working electrode and the counter electrode as well as the detecting electrode after the electrical change is detected in the first change step; and a current measuring step of measuring a current generated between the working electrode and the counter electrode as well as the detecting electrode, to which the voltage is applied in the second application step.
According to of the present invention, there is provided a quantification method for quantifying, by employing the biosensor, a substrate included in a sample liquid supplied to the biosensor comprising: a third application step of applying a voltage between the detecting electrode and the counter electrode or the working electrode as well as between the working electrode and the counter electrode; a reagent supplying step of supplying the sample liquid to the reagent layer; a first change detecting step of detecting an electrical change occurring between the detecting electrode and the counter electrode or the working electrode by the supply of the sample liquid to the reagent layer; a second change detecting step of detecting an electrical change occurring between the working electrode and the counter electrode by the supply of the sample liquid to the reagent layer; a second application step of applying a voltage between the working electrode and the counter electrode as well as the detecting electrode after the electrical changes are detected in the first change detecting step and the second change detecting step; and a current measuring step of measuring a current generated between the working electrode and the counter electrode as well as the detecting electrode, to which the voltage is applied in the second application step.
According to of the present invention, in the quantification method, the second change detecting step is followed by a no-change informing step of informing a user that no change occurs when it is detected that no electrical change occurs between the detecting electrode and the counter electrode or the working electrode for a prescribed period of time.
According to of the present invention, there is provided a quantification apparatus, to which the biosensor is detachably connected and which quantifies a substrate included in a sample liquid supplied to the biosensor comprising: a first current/voltage conversion circuit for converting a current from the working electrode included in the biosensor into a voltage; a first A/D conversion circuit for digitally converting the voltage from the current/voltage conversion circuit; a first switch provided between the counter electrode included in the biosensor and the ground; and a control part for controlling the fist A/D conversion circuit and the first switch, and the control part applies a voltage between the detecting electrode and the working electrode in a state where the first switch is insulated from the counter electrode, detects an electrical change between the detecting electrode and the working electrode occurring by the sample liquid which is supplied to the reagent layer on the specimen supply path, thereafter applies a voltage between the working electrode and the counter electrode as well as the detecting electrode in a state where the first switch is connected to the counter electrode, and measures a response current generated by applying the voltage.
According to of the present invention, there is provided a quantification apparatus, to which the biosensor is detachably connected and which quantifies a substrate included in a sample liquid supplied to the biosensor comprising: a first current/voltage conversion circuit for converting a current from the working electrode included in the biosensor into a voltage; a second current/voltage conversion circuit for converting a current from the detecting electrode included in the biosensor into a voltage; a first A/D conversion circuit for digitally converting the voltage from the first current/voltage conversion circuit; a second A/D conversion circuit for digitally converting the voltage from the second current/voltage conversion circuit; a first selector switch for switching the connection of the detecting electrode of the biosensor to the first current/voltage conversion circuit or the ground; and a control part for controlling the fist A/D conversion circuit, the second A/D conversion circuit, and the first selector switch, and the control part applies a voltage between the detecting electrode and the counter electrode as well as between the working electrode and the counter electrode in a state where the first selector switch is connected to the first current/voltage conversion circuit, detects an electrical change between the detecting electrode and the working electrode as well as an electrical change between the working electrode and the counter electrode, respectively, occurring by the sample liquid which is supplied to the reagent layer provided on the specimen supply path, thereafter connects the first selector switch to the ground, applies a voltage between the working electrode and the counter electrode as well as the detecting electrode, and measures a response current generated by applying the voltage.
According to of the present invention, the quantification apparatus comprises: a second selector switch for switching the connection of the working electrode of the biosensor to the second current/voltage conversion circuit or the ground, and the control part applies a voltage between the detecting electrode and the counter electrode as well as between the working electrode and the counter electrode in a state where the first selector switch is connected to the first current/voltage conversion circuit and the second selector switch is connected to the second current/voltage conversion circuit, respectively, connects the second selector switch to the ground when detecting an electrical change between the working electrode and the counter electrode, occurring by the sample liquid which is supplied to the reagent layer provided on the specimen supply path, and when thereafter detecting an electrical change between the detecting electrode and the working electrode, in a state where the second selector switch is connected to the second current/voltage conversion circuit and the first selector switch is connected to the ground, applies a voltage between the working electrode and the counter electrode as well as the detecting electrode, and measures a response current generated by applying the voltage.
According to of the present invention, the quantification apparatus comprising an informing means for informing a user that no change occurs, when the sample liquid is supplied to the reagent layer of the specimen supply path, and the control part detects that an electrical change occurs between the working electrode and the counter electrode but no electrical change occurs between the detecting electrode and the working electrode or the counter electrode.
BRIEF DESCRIPTION OF DRAWINGS FIG. 1 are exploded perspective views of a biosensor according to a first and a fifth embodiments.
FIG. 21 are exploded perspective views of a conventional biosensor.
FIG. 24 are top views illustrating states of electrodes of a biosensor in a manufacturing method according to the third embodiment.
BEST MODE TO EXECUTE THE INVENTION Hereinafter, embodiments of the present invention will be described with reference to the figures. The embodiments which are described here are merely examples, and the present invention is not necessarily restricted thereto.
Embodiment 1 A biosensor A as defined in Claims 1 to 10 of the present invention will be described as a first embodiment with reference to the figures.
Numeral 1 denotes a first insulating support (hereinafter, referred to as merely �support�) composed of polyethylene terephthalate or the like. Numeral 2 denotes a conductive layer which is formed on the whole surface of the support 1 and composed of an electrical conductive material such as a noble metal, for example gold or palladium, and carbon. Numerals 3 a and 3 b denote slits which are provided on the conductive layer 2 on the support 1 and are parallel to the side of the support 1. Numerals 4 a and 4 b denote slits which are provided on the conductive layer 2 on the support 1 and are vertical to the side of the support 1. Numerals 5, 6, and 7 denote a working electrode, a counter electrode, and a detecting electrode, which are formed by dividing the conductive layer 2 by the slits 3 a and 3 b, as well as 4 a and 4 b. Numeral 8 denotes a spacer which covers the working electrode 5, the counter electrode 6, and the detecting electrode 7 on the support 1. Numeral 9 denotes a rectangular cutout part provided in the middle of an entering edge part of the spacer 8 to form a specimen supply path. Numeral 9 a denotes an inlet of the specimen supply path, numeral 10 denotes a longitudinal width of the cutout part 9 of the spacer 8, and numeral 11 denotes an clearance between the two slits 4 a and 4 b which are provided on the conductive layer 2. Numeral 12 denotes a reagent layer which is formed by applying a reagent including enzyme or the like to the working electrode 5, the counter electrode 6, and the detecting electrode 7 which are exposed from the cutout part 9 of the spacer 8. Numeral 13 denotes a cover (second insulating support) for covering the spacer 8, and numeral 13 a denotes an air hole provided in the middle of the cover 13.
Embodiment 2 A biosensor B according to Claims 11 and 12 of the present invention will be described as a second embodiment.
FIG. 3 are perspective views illustrating the biosensor B in the order of the manufacturing process, and FIG. 4 is a diagram illustrating a specimen supply path of the biosensor B.
Embodiment 3 A specific method for manufacturing the above-described biosensors A and B will be further described. Here, the biosensors A and B are assumed a biosensor X collectively.
Embodiment 4 A biosensor D according to Claims 15 and 16 of the present invention will be described as a fourth embodiment.
FIG. 9 are perspective views illustrating the biosensor D in the order of a manufacturing process. FIG. 10 are top views exemplifying the formation of fourth slits of the biosensor D. FIG. 22 is a diagram illustrating a state where the biosensor D is inserted into a measuring device.
Next, as shown in FIG. 9( b), the first slits 63 a, 63 b, 63 c, and 63 d are formed on the electrical conductive layer 62 by employing the laser, to divide the electrical conductive layer 62 into the working electrode 65, the counter electrode 66, and the detecting electrode 67. Further, the fourth slits 64 a, 64 b, and 64 c are formed on the electrodes, i.e., the working electrode 65, the counter electrode 65, and the detecting electrode 67 by employing the laser. Here, the fourth slits 64 a, 64 b, and 64 c divide all the electrodes, i.e., the working electrode 65, the counter electrode 66, and the detecting electrode 67, while there are for example eight kinds of combinations possible as shown in FIG. 10 as the manner in which the fourth slits 64 a, 64 b, and 64 c are provided.
The combinations of the fourth slits 64 a, 64 b, and 64 c enable the measuring device 4115 to discriminate information of correction data for correcting a difference in the output characteristics for each production lot. For example, in the case of FIG. 10( a) where no fourth slit is provided, it is assumed a biosensor which has output characteristics of the production lot number �1�. In the case of FIG. 10( b) where the fourth slit 64 a is provided only in the counter electrode 66, it is assumed a biosensor which has output characteristics of the production lot number �2�.
Also, the measuring device 4115 checks whether the respective electrodes of the biosensor D, that is, the working electrode 65, the counter electrode 66, and the detecting electrode 67 are divided by the fourth slits 64 a, 64 a, and 64 b. For example, when the electrical conduction between the measuring part 71 and the correction part 57 is checked, it can be seen whether the fourth slit 64 c has been formed. Similarly, when electrical conduction between the measuring part 72 and the correction part 58 is checked, it can be seen whether the fourth slit 64 a has been formed, and when electrical conduction between the measuring part 73 and the correction part 59 is checked, it can be seen whether the fourth slit 64 b has been formed. For example, when the fourth slit is not formed on any electrodes, it is in a state shown in FIG. 10( a) where the biosensor is of the production lot number �1�, and thus the measuring device 4115 obtains a blood sugar level on the basis of the correction data corresponding to the output characteristics of the production lot number �1� which are previously stored and the measured current value, and displays the blood sugar level at the display part 4117. Similarly, when the fourth slit 64 a is formed only in the counter electrode 66, a blood sugar level is obtained on the basis of the correction data corresponding to the output characteristics of the production lot number �2� and the measured current value, and the obtained blood sugar level is displayed at the display part 4117.
In any of the above-described biosensors A, B, C, and D according to the first to fourth embodiments, it is more preferable that each slit provided on the electrical conductive layer is processed by the laser, the width of each slit is 0.005 mm-0.3 mm, and the depth of each slit is equal to or larger than the thickness of the electrical conductive layer
Embodiment 5 A thin film electrode forming method of the present invention will be described as a fifth embodiment with reference to the figures. When the thin film electrode method described in the fifth embodiment is applied when the electrode parts of any of the biosensors A, B, C, and D according to the above-described first to fourth embodiments are formed, a biosensor of the present invention can be obtained.
FIG. 11 is a schematic diagram showing a state of a biosensor, where a thin film electrode is formed by implementing the thin film electrode forming method according to this embodiment and a reaction reagent layer are laid out thereon. This biosensor differs most from the conventional biosensor shown in FIG. 25 in that a surface roughening processing is performed on the surface of an insulating resin support 81 of polyethylene terephthalate, polycarbonate or the like, to enhance adhesion between the support 81 and an electrode layer 82 as well as between the electrode layer 82 and a reaction reagent layer 83. It also differs in that a material constituting the electrode layer 82 is a simple substrate material composed of a noble metal or carbon, and the thickness of the electrode layer 82 is controlled within 3-100 nm.
Embodiment 6 Hereinafter, a quantification method of quantifying a substrate and a quantification apparatus for quantifying a substrate of the present invention, which employ any of the biosensors A, B, C, and D, for which the electrical conductive layers are formed by employing the above-described thin film electrode forming method according to the fifth embodiment will be described. While the biosensor A as described in the first embodiment is used as a biosensor employed in a following description, the biosensor to be used is not restricted thereto.
In the quantification apparatus M1, numerals 115 a, 115 b, and 115 c denote connectors connected to a working electrode 5, a detecting electrode 7, a counter electrode 6 of the biosensor A, respectively, numeral 116 a denotes a switch provided between the connector 115 c and the ground (which means a constant potential electrodeposition and can be not always �0�. The same goes for in the present specification.), numeral 118 a denotes a current/voltage conversion circuit which is connected to the connector 115 a and converts a current flowing between the working electrode 6 and other electrode into a voltage to be output, numeral 119 a denotes an A/D conversion circuit which is connected to the current/voltage conversion circuit 118 a and converts a voltage value from the current/voltage conversion circuit 118 a into a pulse, numeral 120 denotes a CPU which controls ON/OFF of the switch 116 a and calculates the amount of a substrate included in a specimen based on the pulse from the A/D conversion circuit 119 a, and numeral 121 denotes a LCD (liquid crystal display) which displays a measured value calculated by the CPU 20.
Embodiment 7 Hereinafter, a quantification method for quantifying a substrate and a quantification apparatus for quantifying a substrate, which employ any of the biosensors A to D whose electrical conductive layers are formed by employing the thin film electrode forming method described in the fifth embodiment but which are different from those of the above-described sixth embodiment will be described. A biosensor which is employed in a following description is supposed to be the biosensor A described in the first embodiment.
Embodiment 8 Hereinafter, a quantification method for quantifying a substrate and a quantification apparatus for quantifying a substrate, which employ any of the biosensors A to D whose electrical conductive layers are formed by employing the thin film electrode forming method described in the fifth embodiment but are different from those of the above-described sixth and seventh embodiments will be described. The biosensor employed in a following description is supposed to be the biosensor A described in the first embodiment.
APPLICABILITY IN INDUSTRY As described above, the biosensor according to the present invention can be formed by a simple manufacturing method, as well as a biosensor which is excellent in a measuring accuracy, a biosensor in which a reagent layer is placed uniformly on electrodes regardless of a reagent liquid composition, resulting in an uniform performance, a biosensor which can keep the performance constant without affecting an area of an electrode when the support is cut, and a biosensor which enables a discrimination of correction data for each production lot only by being inserted without a correction chip inserted can be obtained, and further the thin film electrode forming method according to the invention is suitable for forming an electrical conductive layer of the biosensor, and further the method and the apparatus for quantification according to the invention are quite useful for diagnostics a slight amount of specimen.
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