Source: http://www.google.com/patents/US20030100030?dq=5,838,906
Timestamp: 2015-01-31 06:42:44
Document Index: 741370963

Matched Legal Cases: ['art 1', 'art 4', 'art 5', 'art 4', 'art 5', 'art 4', 'art 5', 'art 4', 'art 5', 'art 1', 'art 1', 'art 5', 'art 4', 'art 5', 'art 4', 'art 5', 'art 4', 'art 5', 'art 1', 'art 1', 'art 8', 'art 4', 'art 4', 'art 5', 'art 3', 'art 5', 'art 4', 'art 5', 'art 5', 'art 5', 'art 5', 'art 5', 'art 5', 'art 5', 'art 4', 'art 5', 'art 4', 'art 5', 'art 4', 'art 5', 'art 4', 'art 5', 'art 4', 'art 5', 'art 1', 'art 1', 'art 4', 'art 4', 'art 4', 'art 5', 'art 3', 'art 5', 'art 5', 'art 5', 'art 5', 'art 5', 'art 4', 'art 5', 'art 4', 'art 5', 'art 9', 'art 4', 'art 5', 'art 4', 'art 5', 'art 8', 'art 9', 'art 1', 'art 1', 'art 8', 'art 4', 'art 4', 'art 4', 'art 5', 'art 3', 'art 5', 'art 5', 'art 5', 'art 5', 'art 5', 'art 5', 'art 4', 'art 5', 'art 5', 'art 5', 'art 5', 'art 4', 'art 5', 'art 4', 'art 5', 'art 5', 'art 8', 'art 4', 'art 5', 'art 4', 'art 5', 'art 8', 'art 9', 'art 2', 'art 1', 'art 2', 'art 3', 'art 1', 'art 10', 'art 1', 'art 10', 'art 5', 'art 1', 'art 1', 'art 8', 'art 4', 'art 4', 'art 5', 'art 10', 'art 10', 'art 1', 'art 10', 'art 1', 'art 10', 'art 1', 'art 10', 'art 5', 'art 5', 'art 5', 'art 5', 'art 5', 'art 5', 'art 5', 'art 4', 'art 5', 'art 4', 'art 5']

Patent US20030100030 - Biosensor for use as diagnostic tool in medicine - Google PatentsSearch Images Maps Play YouTube News Gmail Drive More »Sign inAdvanced Patent SearchPatentsA biosensor according to the present invention includes a reagent immobilization part (5) where an antibody for a measurement target in an inspection target solution is immobilized, and a marked reagent holding part (4) where an antibody which is marked at a part of a development layer in a dry state...http://www.google.com/patents/US20030100030?utm_source=gb-gplus-sharePatent US20030100030 - Biosensor for use as diagnostic tool in medicineAdvanced Patent SearchPublication numberUS20030100030 A1Publication typeApplicationApplication numberUS 10/048,727PCT numberPCT/JP2001/004498Publication dateMay 29, 2003Filing dateMay 29, 2001Priority dateMay 29, 2000Also published asCN1189751C, CN1380977A, DE60111904D1, DE60111904T2, EP1202060A1, EP1202060A4, EP1202060B1, WO2001092884A1Publication number048727, 10048727, PCT/2001/4498, PCT/JP/1/004498, PCT/JP/1/04498, PCT/JP/2001/004498, PCT/JP/2001/04498, PCT/JP1/004498, PCT/JP1/04498, PCT/JP1004498, PCT/JP104498, PCT/JP2001/004498, PCT/JP2001/04498, PCT/JP2001004498, PCT/JP200104498, US 2003/0100030 A1, US 2003/100030 A1, US 20030100030 A1, US 20030100030A1, US 2003100030 A1, US 2003100030A1, US-A1-20030100030, US-A1-2003100030, US2003/0100030A1, US2003/100030A1, US20030100030 A1, US20030100030A1, US2003100030 A1, US2003100030A1InventorsMasakata Nadaoka, Mie Takahashi, Hirotaka Tanaka, Fumihisa KitawakiOriginal AssigneeNadaoka Masakata ., Mie Takahashi, Hirotaka Tanaka, Fumihisa KitawakiExport CitationBiBTeX, EndNote, RefManReferenced by (5), Classifications (11), Legal Events (3) External Links: USPTO, USPTO Assignment, EspacenetBiosensor for use as diagnostic tool in medicineUS 20030100030 A1Abstract A biosensor according to the present invention includes a reagent immobilization part (5) where an antibody for a measurement target in an inspection target solution is immobilized, and a marked reagent holding part (4) where an antibody which is marked at a part of a development layer in a dry state to be eluted by development of the inspection target solution is held, at parts of the development layer for developing the inspection target solution, and measures a bonding amount of the marked reagent bonded in the reagent immobilization part (5), thereby qualitatively and quantitatively measuring a measurement component in the inspection target solution. In such bionsensor, side faces of the development layer which are parallel to the direction of the inspection target solution permeating are partially or entirely melted and cured to be sealed. According to the so-constituted biosensor, the permeation of the inspection target solution in the development layer is arranged, and a low-cost and high-accuracy biosensor and a manufacturing method thereof can be realized by a simplified biosensor manufacturing method. Images(10) Claims(21)
BRIEF DESCRIPTION OF DRAWINGS [0058]FIG. 1 are perspective views illustrating constitutions of a biosensor according to a first embodiment of the present invention. [0059]FIG. 2 are perspective views illustrating constitutions of a biosensor according to a second embodiment of the present invention. [0060]FIG. 3 are perspective views illustrating constitutions of a biosensor according to third embodiment of the present invention. [0061]FIG. 4 are diagrams illustrating constitutions of a cross section of a biosensor before and after cutting, in which a reagent holding part is not in contact with parallel side faces of a development layer according to a fourth embodiment of the present invention. [0062]FIG. 5 are diagrams illustrating constitutions of a cross section of a biosensor before and after cutting, in which the reagent holding part is in contact with the parallel side faces of the development layer according to the fourth embodiment of the invention. [0063]FIG. 6 illustrates a micrograph of a cross section of a biosensor which is cut in a conventional cutting process. [0064]FIG. 7 illustrates a micrograph of a cross section of a biosensor which is cut in a cutting process according to the fourth embodiment of the invention. [0065]FIG. 8 are perspective views illustrating constitutions of a biosensor according to a fifth embodiment of the present invention. [0066]FIG. 9 are pattern diagrams illustrating measurement states of the biosensor according to the fifth embodiment of the invention. [0067]FIG. 10 are graphs illustrating relationships between hCG concentration and a measurement result in two kinds of immuno-chromatographic specimens according to an example of the present invention.
BEST MODE TO EXECUTE THE INVENTION [0068] Hereinafter, embodiments of the present invention will be described. [0069] (Embodiment 1) [0070] A biosensor and a manufacturing method thereof according to a first embodiment of the present invention make an inspection target solution permeate on a development layer at a uniform rate. [0071] First, the constitution of the biosensor according to the first embodiment will be described with reference to FIG. 1. [0072]FIG. 1 are perspectives views illustrating constitutions of the biosensor which is sealed by melting and curing; FIG. 1(a) illustrates the biosensor, which is constituted only by the development layer, FIG. 1(b) illustrates the biosensor, which is provided with a liquid-impermeable sheet material on the development layer, and FIG. 1(c) illustrates the biosensor, which is provided with the liquid-impermeable sheet material on the development layer and a base material thereunder. [0073] In FIG. 1, numeral 1 denotes an application part 1 for applying an inspection target solution onto the development layer, which is composed of a nonwoven fabric. Numeral 2 denotes a reaction part in the reactive layer, which is composed of a nitrocellulose. Numeral 3 denotes a water absorbing part for absorbing the solution permeating on the development layer, which is composed of glass fiber filter paper. These materials used for the development layer may be arbitrary porous materials which can be permeated by the inspection target solution, such as filter paper, a nonwoven fabric, a membrane, a fabric or the like. [0074] Numeral 4 denotes a marked reagent holding part at a part on the development layer, in which an antibody for a measurement target in the inspection target solution is held, being marked with a gold colloid, in a dry state so as to be eluted by the inspection target solution. Numeral 5 denotes a reagent immobilization part, in which the antibody for the measurement target in the inspection target solution is immobilized in a dry state so as to be bonded with a different epitope as that for the marked reagent to form a complex of the immobilized antibody, the measurement target in the inspection target solution and the marked reagent. In the first embodiment, the marked reagent holding part 4 and the reagent immobilization part 5 are formed in spot shapes at parts on the development layer, and particularly constituted to prevent the reagent from being in contact with side faces which are parallel to the direction of the inspection target solution permeating. While the marked reagent holding part 4 and the reagent immobilization part 5 have the spot shapes, they do not necessarily have the spot shapes, and any shapes can be selected freely as long as the marked reagent holding part 4 and the reagent immobilization part 5 are formed in shapes which prevent their contacts with the parallel side faces of the development layer. Further, while the development layer having the marked reagent holding part 4 and the reagent immobilization part 5 is constituted to cause a so-called sandwich reaction in the antigen antibody reaction, the development layer may be constituted to cause a competitive reaction when a reagent which competitively reacts to the measurement target in the inspection target solution is employed according to selection of the reagent. Further, when a specific bonding other than a bonding caused by the antigen antibody reaction is to be utilized, the development layer can be constituted by an arbitrary reagent component. With respect to a marking method, the above-described gold colloid is only an example, and others, such as an enzyme, a protein, coloring matters, a fluorescene, a metallic sol, a nonmetallic sol, and coloring particles such as a latex, can be selected arbitrarily according to need. [0075] Numeral 6 denotes a liquid-permeable sheet material, which is composed of transparent PET tape. This liquid-permeable sheet material 6 adherently covers the development layer except for the application part 1. The liquid-permeable sheet material 6 covers the above-described part of the development layer, thereby protectingly intercepting application of the inspection target solution to parts other than the application part 1 as well as preventing external pollution due to the inspection target solution being improvidently in contact with the development layer or an inspector directly touching the development layer with his/her hand or the like. It is preferred that a transparent material is employed for this liquid-permeable sheet material 6, and particularly a part for covering the reagent immobilization part 5, which is a part for confirming a measurement result, is at least in a permeable state. In a case where the biosensor does not require a high-accuracy result or the formed development layer is finally put in a hollow casing, the liquid-permeable sheet material 6 is not indispensable. [0076] Numeral 7 denotes a base material for holding the development layer, which is composed of a white PET film, for example. The base material 7 reinforces the development layer, and when a solution which poses a risk of infection, such as blood, saliva, and urine, is employed as the inspection target solution, the base material 7 intercepts it. Further, it is also possible that the base material 7 intercepts light in a case where the development layer is permeated and becomes light permeable. While the base material 7 is arranged under the development layer here, the liquid-permeable sheet material 6, instead of the base material 7, may cover the rear face of the development layer. When the solution which poses a risk of infection, such as blood, saliva, and urine, is employed as the inspection target solution, that solution is intercepted also in this case, and the number of materials of the biosensor can be decreased, thereby further reducing the cost. [0077] Numeral 8 denotes a melted and cured part, which is formed by melting the parallel side faces of the development layer by a CO2 laser and thereafter curing the same by cooling. Since in the first embodiment the marked reagent holding part 4 and the reagent immobilization part 5 are not in contact with the parallel side faces of the development layer, the marked reagent holding part 4 and the reagent immobilization part 5 are not affected even when the side faces thereof are melted. Therefore, the amount of the reagent in the marked reagent holding part 4 and the reagent immobilization part 5 can be minimized, and this is effective when the reagent is expensive or minute in amount. Further, while in the first embodiment the CO2 laser is employed as a melting method, others such as an excimer laser, an argon laser, a YAG laser, a helium neon laser, a ruby laser and the like can be also employed. In addition to the laser, the parallel side faces of the development layer can be also melted by bringing a heated metal or the like into contact therewith, as well as by an organic solvent or the like depending on a material of the development layer. The above-described melting methods are only examples, and an arbitrary method is available as long as the parallel side faces of the development layer can be melted. [0078] Next, a measuring method of the so-constituted biosensor according to the first embodiment will be described with reference to FIG. 1. The biosensor exemplified in the first embodiment is a specimen for a one-step immuno-chromatography, and a basic measurement operation using the biosensor comprises only one operation of applying the inspection target solution so as to start a measurement. The immuno-chromatography here is an immunoassay method in which a permeable porous material is employed and a complex of the immobilized reagent and the marked reagent is formed, thereby performing a measurement, and is a system of measurement that utilizes the antigen antibody reaction, in which B/F separation is implemented in the process of the inspection target solution permeating a chromatography carrier, while a washing operation such as the B/F separation is required in a usual immunoassay method. In this immuno-chromatography, whole reagent is usually in a dry state, and a marker for marking the reagent is usually a gold colloid or a latex, while magnetic particles, an enzyme, a metallic colloid or the like are also used. When the marker is an enzyme or the like, there is included an operation of adding an enzyme substrate or a reaction stop reagent as a measurement operation by a user. [0079] A measurement by the so-constituted biosensor starts when an appropriate amount of inspection target solution is applied to the application part 1. When the inspection target solution is applied to the application part 1, the inspection target solution permeates the development layer. Since the development layer has its side faces sealed with the melted and cured part 8, a permeation rate of the inspection target solution at the side faces is not increased, resulting in permeation at a uniform rate. Then, when the inspection target solution reaches to the marked reagent holding part 4, the marked reagent held in the marked reagent holding part 4 starts to be eluted. Thereafter, the permeation of the inspection target solution being promoted, the inspection target solution reaches to the reagent immobilization part 5 and then absorbed in the water absorbing part 3. [0080] Then, the measurement result is obtained by confirming a bonding state of the marked reagent in the reagent immobilization part 5 in a predetermined period of time. Since in the first embodiment the marked reagent holding part 4 and the reagent immobilization part 5 are not in contact with the parallel side faces of the development layer and the parallel side faces are melted to be sealed, uniform permeation of the inspection target solution is performed in the reagent immobilization part 5, whereby the marked reagent and the measurement target in the inspection target solution, which pass through the reagent immobilization part 5, permeate respective surfaces of the reagent immobilization part 5 uniformly, and thus a partial difference in the bonding amount is eliminated, so that a highly accurate measurement result can be obtained. [0081] Further, the bonding state of the marked reagent in the reagent immobilization part 5 can be measured visually when a qualitative judgement is required. When a more accurate measurement is required, the bonding amount of the marked reagent is measured employing an optical method, thereby obtaining a quantitative result. Further, since in the first embodiment the reagent immobilization part 5 can be shaped so that an inspector easily confirms the result, a visual judgement or the like is suitable as a method of measuring the boding state of the marked reagent in the reagent immobilization part 5. [0082] As described above, according to the biosensor and the manufacturing method thereof in the first embodiment, the marked reagent holding part 4 and the reagent immobilization part 5 are formed in spot shapes at parts on the development layer so as not to be in contact with its parallel side faces, and the parallel side faces are partially or entirely malted and cured so as to be sealed in the direction of the inspection target solution on the development layer permeating, whereby the permeation of the inspection target solution in the development layer can be arranged, resulting in a more accurate quantitative measurement. Further, since the development layer itself is melted and cured, no new material is required for sealing the parallel side faces, and a manufacturing process can be simplified, thereby reducing the cost due to reduction in material and manufacturing process. [0083] Further, a nitrocellulose, which makes bonding of a protein or the like relatively easy, is employed as a material of the development layer, thereby making it easy to manufacture the biosensor. [0084] Further, the parallel side faces of the development layer are melted by a laser and cured so as to be sealed, whereby a required part of the parallel side faces can be melted and cured more speedily and uniformly and the melted and cured part can be given uniformity. Further, the side faces of the development layer can be melted and cured in a no-physical-contact state, so that the shape of the development layer is not changed due to contact pressure, resulting in a high-accuracy measurement. [0085] While in the first embodiment the biosensor utilizing the antigen antibody reaction has been given as an example, any specific bonding reactions are similarly available as well as the antigen antibody reaction. In addition, the measuring operation is not restricted to one which comprises one-step operation of applying the inspection target solution so as to perform a measurement, and one which requires plural operations, such as applying a reaction stop reagent or the like when the marker is an enzyme, or diluting the specimen, in addition to applying the inspection target solution, is also available. [0086] (Embodiment 2) [0087] In a biosensor and a manufacturing method thereof according to a second embodiment of the present invention, a reagent component on side faces parallel to the direction in which an inspection target solution permeates on the development layer, in the marked reagent holding part 4 and the reagent immobilization part 5 is denatured to be deactivated. [0088] First, the constitution of the biosensor according to the second embodiment will be described with reference to FIG. 2. [0089]FIG. 2 are perspective views illustrating the biosensor in a state where the reagent component on the parallel side faces of the development layer are denatured to be deactivated; FIG. 2(a) illustrates the biosensor, which is constituted only by the development layer, FIG. 2(b) illustrates the biosensor, which is provided with a liquid-impermeable sheet material on the development layer, and FIG. 2(c) illustrates the biosensor, which is provided with the liquid-impermeable sheet material on the development layer and a base material thereunder. [0090] In FIG. 2, the biosensor according to the second embodiment is a specimen for the one-step immuno-chromatography as in the first embodiment, in which the marked reagent holding part 4 and the reagent immobilization part 5 are formed on the development layer. In this embodiment, the marked reagent holding part 4 and the reagent immobilization part 5 are formed in strip shapes and the reagent is in contact with the side faces parallel to the direction in which the inspection target solution permeates on the development layer. Numeral 9 denotes a reagent deactivation part, which is obtained by deactivating side faces of the development layer in the marked reagent holding part 4 and in the reagent immobilization part 5 by a CO2 laser. To deactivate the reagent component on the side faces of the development layer, a method of uniformly deactivating the vicinity of the side faces, such as a method of deactivating by bringing a heated metal surface into contact with the side faces or a method of applying and atomizing a solution of acid, alkali or the like which denatures a protein, is preferred. In FIG. 2, the same parts as those shown in FIG. 1 are denoted by the same reference numerals and their descriptions will be omitted. [0091] Next, a measuring method of the so-constituted biosensor according to the second embodiment will be described with reference to FIG. 2. [0092] First, a measurement by the above-described biosensor starts when an appropriate amount of inspection target solution is applied to the application part 1. When the inspection target solution is applied to the application part 1, the inspection target solution permeates on the development layer. When the inspection target solution permeates the marked reagent holding part 4, the marked reagent starts to be eluted. However, the marked reagent on the side faces of the development layer in the marked reagent holding part 4 is deactivated, and therefore that marked reagent is not eluted or has lost its specific property. The specific property here is for example a specific bonding reaction such as the antigen antibody reaction or a specific reaction in the case of employing an enzyme or the like as the marked reagent in the marked reagent holding part 4. [0093] Subsequently, the permeation of the inspection target solution in the development layer being promoted, the inspection target solution reaches to the reagent immobilization part 5 and then absorbed in the water absorbing part 3. Also in this reagent immobilization part 5, the reagent on the side faces of the development layer is denatured to be deactivated, and thus no reaction is caused on the side faces of the reagent immobilization part 5. [0094] The permeation of the inspection target solution in the development layer employs no mechanical operation and thus is performed according to the property of the development layer. Considering that fine pores in the development layer are cut into pieces on the side faces of the development layer and their shapes are different from those at the center part, a permeation rate varies between on the both side faces of the development layer and at the center part. Therefore, in the second embodiment, the reagent on the side faces of the development layer is denatured to be deactivated so that no reaction is caused on the side faces of the development layer as described above, thereby eliminating a bonding of the marked reagent on the side faces of the reagent immobilization part 5 where the permeation of the inspection target solution is ununiform. There is a large number of fine pores here, and the center part of the development layer indicates parts other than the side faces. [0095] Then, the measurement result of the above-described biosensor is obtained by confirming a bonding state of the marked reagent in the reagent immobilization part 5 in a predetermined period of time. The bonding state of the marked reagent in this reagent immobilization part 5 can be measured visually when a qualitative judgement is required. When a more accurate measurement is required, the bonding amount of the marked reagent is measured employing an optical method, thereby obtaining a quantitative result. [0096] As described above, according to the biosensor and the manufacturing method thereof in the second embodiment, the marked reagent holding part 4 and the reagent immobilization part 5 are formed on the development layer in strip shapes, and the parallel side faces of the marked reagent holding part 4 and reagent immobilization part 5 on the development layer are denatured to be deactivated, so that no reaction is caused on the side faces of the development layer, thereby preventing deterioration in measurement accuracy due to ununiform permeation of the inspection target solution. [0097] Further, the reagent deactivation part 9 of the development layer is obtained by denaturing and deactivating the reagent component in no contact therewith by laser irradiation, whereby the shape of the development layer is not changed due to contact pressure in the denaturing and deactivating process, so that ununiform permeation due to the change in the shape of the development layer shape can be prevented, resulting in manufacture of high-accuracy biosensor. [0098] (Embodiment 3) [0099] A biosensor and a manufacturing method thereof according to a third embodiment of the present invention make an inspection target solution permeate on a development layer at a uniform rate, in which side faces of the development layer parallel to the direction of the inspection target solution permeating are melted and cured to be sealed, and at the same time, a reagent component on the parallel side faces of the marked reagent holding part 4 and the reagent immobilization part 5 which are formed on the development layer in strip shapes is denatured to be deactivated. [0100] First, the constitution of the biosensor according to the third embodiment will be described with reference to FIG. 3. [0101]FIG. 3 are perspectives views illustrating constitutions of the biosensor in which parallel side faces of the development layer are sealed by melting and curing and the reagent component thereon is denatured to be deactivated; FIG. 3(a) illustrates the biosensor, which is constituted only by the development layer, FIG. 3(b) illustrates the biosensor, which is provided with a liquid-impermeable sheet material on the development layer, and FIG. 3(c) illustrates the biosensor, which is provided with the liquid-impermeable sheet material on the development layer and a base material thereunder. [0102] In FIG. 3, in the biosensor according to the third embodiment, the marked reagent holding part 4 and the reagent immobilization part 5 are formed in strip shapes as in the second embodiment, and the side faces parallel to the direction in which the inspection target solution is applied on the development layer are entirely melted by irradiating a CO2 laser or the like and cured so as to be sealed (melted and cured part 8), and at the same time, the reagent component on the parallel side faces is denatured to be deactivated (reagent deactivation part 9). In FIG. 3, the same parts as those shown in FIG. 2 are denoted by the same reference numerals and their descriptions will be omitted. [0103] Next, a measuring method of the so-constituted biosensor according to the third embodiment will be described with reference to FIG. 3. [0104] First, a measurement by the above-described biosensor starts when an appropriate amount of inspection target solution is applied to the application part 1. When the inspection target solution is applied to the application part 1, the inspection target solution permeates on the development layer. Since the development layer has its side faces sealed with the melted and cured part 8, a permeation rate at the side faces is not increased, resulting in permeation at a uniform rate. Then, when the inspection target solution permeates the marked reagent holding part 4, the marked reagent starts to be eluted. However, the marked reagent on the side faces of the development layer in the marked reagent holding part 4 is deactivated, and therefore that marked reagent is not eluted or has lost its specific property. The specific property here is for example a specific bonding reaction such as the antigen antibody reaction or a specific reaction in the case of employing an enzyme or the like as the marked reagent in the marked reagent holding part 4. [0105] Subsequently, the permeation of the inspection target solution in the development layer being promoted, the inspection target solution reaches to the reagent immobilization part 5 and then absorbed in the water absorbing part 3. Also in this reagent immobilization part 5, the reagent on the side faces of the development layer is denatured to be deactivated, and thus no reaction is caused on the side faces of the reagent immobilization part 5. [0106] The measurement result of this biosensor can be obtained by confirming a bonding state of the marked reagent in the reagent immobilization part 5 in a predetermined period of time. Since in the third embodiment the parallel side faces of the development layer are melted to be sealed, uniform permeation of the inspection target solution is performed in the reagent immobilization part 5, whereby the marked reagent and the measurement target in the inspection target solution, which pass through the reagent immobilization part 5, permeate respective surfaces of the reagent immobilization part 5 uniformly, and thus a partial difference in the bonding amount is eliminated, resulting in a highly accurate measurement result. Further, in the third embodiment, the parallel side faces of the development layer in the marked reagent holding part 4 and the reagent immobilization part 5 are denatured to be deactivated, whereby ununiform permeation of the inspection target solution in the reagent immobilization part 5 which is due to variation in a permeation rate between on the side faces of the development layer and at the center part can be eliminated by preventing the bonding of the marked reagent on the side faces of the reagent immobilization part 5, resulting in a more accurate and uniform measurement result. The center part of the development layer here indicates parts other than the side faces. Further, the bonding state of the marked reagent in the reagent immobilization part 5 can be measured visually when a qualitative judgement is required. When a more accurate measurement is required, the bonding amount of the marked reagent is measured employing an optical method, thereby obtaining a quantitative result. [0107] As described above, according to the biosensor and the manufacturing method thereof in the third embodiment, the marked reagent holding part 4 and the reagent immobilization part 5 are formed on the development layer in strip shapes, and the side faces parallel to the direction in which the inspection target solution permeates on the development layer are partially or entirely melted and cured so as to be sealed, and at the same time, the reagent component on the parallel side faces in the marked reagent holding part 4 and the reagent immobilization part 5 is denatured to be deactivated, whereby the permeation of the inspection target solution into the development layer is arranged and deterioration in accuracy due to the permeation into the parallel side faces in the reagent immobilization part 5 is eliminated, resulting in a more accurate quantitative measurement. Further, since the development layer itself is melted and cured, no new material is required for sealing the parallel side faces, and a manufacturing process can be simplified, thereby reducing the cost due to reduction in material and manufacturing process. [0108] Further, a nitrocellulose, which makes bonding of a protein or the like relatively easy, is employed as a material of the development layer, thereby making it easy to manufacture the biosensor. [0109] Further, the parallel side faces of the development layer are melted by a laser and cured so as to be sealed, whereby a required part of the parallel side faces can be melted and cured more speedily and uniformly and the melted and cured part can be given uniformity. Further, the side faces of the development layer can be melted and cured in a no-physical-contact state, so that the shape of the development layer is not changed due to contact pressure, resulting in a high-accuracy measurement. [0110] (Embodiment 4) [0111] In a biosensor and a manufacturing method thereof according to a fourth embodiment of the present invention, the biosensor is cut and its cut surfaces are sealed in a single process at manufacture. [0112] Hereinafter, a manufacturing process of the biosensor according to the fourth embodiment will be described with reference to FIGS. 4 and 5. [0113]FIG. 4 are A-A′ cross section diagrams before and after the cutting process of the biosensor that has a spot-shaped reagent immobilization part according to the fourth embodiment; FIG. 4(a) illustrates a state before cutting and FIG. 4(b) illustrates a state after cutting. FIG. 5 are A-A′ cross section diagrams before and after the cutting process of the biosensor that has a strip-shaped reagent immobilization part according to the fourth embodiment; FIG. 5(a) illustrates a state before cutting and FIG. 5(b) illustrates a state after cutting. FIGS. 4 and 5 illustrate A-A′ cross sections in FIG. 1(c) and FIG. 3(c), and the same parts as those shown in FIGS. 1 and 3 are denoted by the same reference numerals and their descriptions will be omitted. [0114] In FIG. 4(a), the biosensor is formed in a sheet shape by the development layer, the liquid-impermeable sheet material 6 and the base material 7. The sheet shape here is a state before cutting where the biosensor continue plurally as shown in FIG. 4(b). In the fourth embodiment, the sheet-shaped biosensor in FIG. 4(a) is cut employing a CO2 laser to manufacture plural biosensors as shown in FIG. 4(b) from one sheet-shaped biosensor. In the manufacturing process of the biosensor according to the fourth embodiment, when the sheet-shaped biosensor is cut, its sides faces are melted and cured to form the melted and cured part 8 at the same time as shown in FIG. 4(b), which means the cutting and the sealing of the cut surfaces can be performed in a single process. [0115] Also in FIG. 5(a), the biosensor is formed in a sheet shape, and the sheet-shaped biosensor in FIG. 5(a) is cut employing a CO2 laser here to manufacture plural biosensors as shown in FIG. 5(b) from one sheet-shaped biosensor. Since, in FIG. 5, the marked reagent holding part 4 and the reagent immobilization part 5 are formed in strip shapes, the cutting and the sealing of the cut surfaces, as well as the denaturation and deactivation of a reagent component on side faces of the marked reagent holding part 4 and the reagent immobilization part 5 are performed at the same time in a single process, which means the melted and cured part 8 and the reagent deactivation part 9 can be formed in the biosensor in a single cutting process. [0116] Here, a comparison will be made between a cross section state of the biosensor according to the manufacturing process of the fourth embodiment and a cross section state of a biosensor according to a conventional manufacturing process with reference to FIGS. 6 and 7. FIG. 6 illustrates a micrograph of the side faces parallel to the direction of the inspection target solution permeating in a case where the biosensor is cut with a conventional cutting method, and FIG. 7 illustrates a micrograph of the side faces parallel to the direction of the inspection target solution permeating in a case where the biosensor is cut by the cutting method according to the fourth embodiment. [0117] The side faces of the biosensor manufactured by the manufacturing process of the fourth embodiment, in which the cutting employing a CO2 laser and the melting and curing of the side faces are performed at the same time, are melted and cured to be sealed as shown in FIG. 7. On the other hand, the side faces of the conventional biosensor manufactured by the conventional manufacturing process are changed in shape as the side faces of the development layer are damaged by a cutting tool brought into contact therewith. [0118] In comparison between FIGS. 6 and 7, it is confirmed that the side faces of the development layer in FIG. 6 is deformed, while the side faces of the development layer in FIG. 7 are kept remaining porous and are melted and cured to be sealed. [0119] As described above, according to the biosensor and the manufacturing method thereof in the fourth embodiment, the sheet-shaped biosensor is cut by a laser and the cut surfaces are melted and cured simultaneously to be sealed, thereby requiring no operation for sealing in a process different from the cutting process in the manufacturing process of the biosensor, resulting in a simple, low-cost, and high-accuracy biosensor. [0120] (Embodiment 5) [0121] In a biosensor and a manufacturing method thereof according to a fifth embodiment of the present invention, a development layer is made of a single member, a water absorbing part or the like is eliminated, and a liquid-permeable sheet material at the beginning and end of the development layer is removed, in order to reduce the cost as well as simplify a manufacturing process with high accuracy and precision. [0122] First, the constitution of the biosensor according to the fifth embodiment will be described with reference to FIG. 8. [0123]FIG. 8 are perspective views illustrating the biosensor which is sealed by melting and curing; FIG. 8(a) illustrates the biosensor, which is provided with the liquid-impermeable sheet material on the development layer, and FIG. 8(b) illustrates the biosensor, which is provided with the liquid-impermeable sheet material on the development layer and a base material thereunder. [0124] In FIG. 8, numeral 1 denotes an application part for applying an inspection target solution onto the development layer, which is made of the same member as that of a reaction part 2. Numeral 2 denotes the reaction part in the development layer, which is composed of a nitrocellulose. Numeral 10 denotes an end open part in the development layer which is not covered with the liquid-permeable sheep material. These materials used for the development layer may be arbitrary porous materials which can be permeated by the inspection target solution, such as filter paper, a nonwoven fabric, a membrane, a fabric or the like. [0125] In the case of aiming at reducing the cost of the biosensor as well as simplifying the manufacturing process thereof, it is preferred that the development layer is composed of a single member and further is made of a thick material since the inspection target solution is required to be developed in the development layer even when the inspection target solution is small in amount (100 μL or less) or is extremely small in amount (10 μL or less). Therefore, a membrane such as a nitrocellulose is preferable to be employed as the material of the development layer. [0126] Numeral 4 denotes a marked reagent holding part at a part on the development layer, in which an antibody for a measurement target in the inspection target solution is held, being marked with a gold colloid, in a dry state so as to be eluted by the inspection target solution. Numeral 5 denotes a reagent immobilization part, in which the antibody for the measurement target in the inspection target solution is immobilized in a dry state so as to be bonded with a different epitope as that for the marked reagent to form a complex of the immobilized antibody, the measurement target in the inspection target solution and the marked reagent. Further, while the development layer is constituted to cause a so-called sandwich reaction in the antigen antibody reaction, a competitive reaction is also available when a reagent which competitively reacts to the measurement target in the inspection target solution is employed according to selection of the reagent. Further, when a specific bonding other than a bonding caused by the antigen antibody reaction is to be utilized, the development layer can be constituted by an arbitrary reagent component. With respect to a marking method, the above-described gold colloid is only an example, and others, such as an enzyme, a protein, coloring matters, a fluorescene, a metallic sol, a nonmetallic sol, and coloring particles such as a latex, can be selected arbitrarily according to need. [0127] In the case of aiming at improving accuracy as well as reducing the cost of the biosensor and simplifying the manufacturing process thereof, it is preferred that the above-described both marked reagents are held or immobilized in the same member, the application part 1, the reaction part 2 and the water absorbing part 3 are not separated members but a single member, and the both reagent components are provided at different parts on the same plane. [0128] Numeral 6 denotes a liquid-permeable sheet material, which is composed of transparent PET tape. This liquid-permeable sheet material 6 adherently covers the development layer except for the application part 1 and the end open part 10 of the development layer. The liquid-permeable sheet material 6 covers the above-described part of the development layer, thereby protectingly intercepting application of the inspection target solution to parts other than the application part 1 and the end open part 10 as well as preventing external pollution due to the inspection target solution being improvidently in contact with the development layer or an inspector directly touching the development layer with his/her hand or the like. It is preferred that a transparent material is employed for this liquid-permeable sheet material 6, and particularly a part for covering the reagent immobilization part 5, which is a part for confirming a measurement result, is at least in a permeable state. Further, it is preferable that the liquid-impermeable sheet material 6 adherently covers the development layer in order to obtain a high-accuracy measurement result by giving uniformity in permeation to the inspection target solution that is permeating the development layer. [0129] Numeral 7 denotes a base material for holding the development layer, which is composed of a white PET film, for example. The base material 7 reinforces the development layer, and when a solution which poses a risk of infection, such as blood, saliva, and urine, is employed as the inspection target solution, the base material 7 intercepts it. Further, it is also possible that the base material 7 intercepts light in a case where the development layer is permeated and becomes light permeable. [0130] Numeral 8 denotes a melted and cured part, which is formed by melting parallel side faces of the development layer by a CO2 laser and thereafter curing the same by cooling. While the CO2 laser is employed as a melting method here, others such as an excimer laser, an argon laser, a YAG laser, a helium neon laser, a ruby laser and the like can be also employed. In addition to the laser, the parallel side faces of the development layer can be also melted by bringing a heated metal or the like into contact therewith, as well as by an organic solvent or the like depending on a material of the development layer. The above-described melting methods are only examples, and an arbitrary method is available as long as the parallel side faces of the development layer can be melted. [0131] Next, a measuring method of the so-constituted biosensor according to the fifth embodiment will be described with reference to FIG. 9. [0132]FIG. 9 are pattern diagrams illustrating measurement states of the biosensor according to the fifth embodiment. [0133] A measurement by the above-described biosensor is started when an appropriate amount of inspection target solution 11 is applied to the application part 1 (FIG. 9(a)). When the inspection target solution 11 is applied to the application part 1, the inspection target solution 11 permeates the development layer (FIG. 9(b)). Since the development layer has its side faces sealed with the melted and cured part 8, a permeation rate of the inspection target solution 11 at the side faces is not increased, resulting in permeation in the development layer at a uniform rate. Then, when the inspection target solution 11 reaches to the marked reagent holding part 4, the marked reagent held in the marked reagent holding part 4 starts to be eluted. Thereafter, the permeation of the inspection target solution 11 being promoted, the inspection target solution 11 reaches to the reagent immobilization part 5 and then to the end open part 10 (FIG. 9(c)). The inspection target solution 11 which reaches to the end open part 10 is dried by evaporation 12 and further exuded to the same height as that of the inspection target solution 11 in the application part 1 in relation to a water head difference (FIG. 9(d)). [0134] Therefore, this biosensor does not require a new material for absorbing extra inspection target solution 11 and enables the inspection target solution 11 to permeate the development layer in the predetermined direction, from the direction of the inspection target solution 1 to the direction of the end open part 10. While in the fourth embodiment the application part 1 and the end open part 10 are exposed and opened, it is also possible according to need that the application part 1 and the end open part 10 are constituted by a protective material or that the development layer with the above-described constitution is put in a hollow casing so that the inspection target solution 11 is not attached to inspector's hand, in a case for example where the inspection target solution 11 is staining substance. [0135] Then, the measurement result is obtained by confirming a bonding state of the marked reagent in the reagent immobilization part 5 in a predetermined period of time. Since in the present invention the parallel side faces of the development layer in the reagent immobilization part 5 are melted to be sealed as described above, uniform permeation of the inspection target solution 11 in the development layer is performed, whereby the marked reagent and the measurement target in the inspection target solution 11, which pass through the reagent immobilization part 5, permeate respective surfaces of the reagent immobilization part 5 uniformly, and thus a difference in the bonding amount of the marked reagent between at the center of the reagent immobilization part 5 and on the side faces is eliminated, resulting in a highly accurate measurement result. [0136] The marked reagent in this reagent immobilization part 5 can be measured visually when a qualitative judgement is required. When a more accurate measurement is required, the bonding amount of the marked reagent in the reagent immobilization part 5 is measured employing an optical method, thereby obtaining a quantitative result. [0137] While in the fifth embodiment a description has been given taking a case where the marked reagent holding part 4 and the reagent immobilization part 5 are formed on the development layer in strip shapes as an example, the marked reagent holding part 4 and the reagent immobilization part 5 may be formed also in spot shapes. [0138] As described above, according to the biosensor and the manufacturing method thereof in the fifth embodiment, the side faces of the development layer parallel to the direction of the inspection target solution 11 permeating are partially or entirely melted and cured to be sealed, so that the permeation of the inspection target solution 11 is arranged, thereby enabling a more accurate quantitative measurement, and further the development layer except for its end and beginning is adherently covered with the liquid-permeable sheet material 6, whereby it is possible to eliminate the use of a member for the part for applying the inspection target solution and a member for the part for absorbing the inspection target solution. [0139] Further, since the development layer itself is melted and cured, no new material is required for sealing the parallel side faces, and a manufacturing process can be simplified, whereby the cost can be reduced due to reduction in material and manufacturing process, while sensor performance is maintained. [0140] Further, a nitrocellulose, which makes bonding of a protein or the like relatively easy, is employed as a material of the development layer, and the reagent component is formed on the same plane as the development layer, thereby making it easy to manufacture the biosensor. [0141] Further, the parallel side faces of the development layer are melted by a laser and cured so as to be sealed, whereby a required part of the parallel side faces can be melted and cured more speedily and uniformly and the melted and cured part can be given uniformity. Further, by the laser melting, the parallel side faces can be melted and cured in no physical contact with the development layer, so that the shape of the development layer is not changed due to contact pressure, resulting in a high-accuracy measurement. EXAMPLE [0142] Hereinafter, the present invention will be described more specifically through an example, while the present invention is not restricted to descriptions in this example in the scope of the invention. [0143] In this embodiment, the immuno-chromatography development layer which includes an anti-hCG-β antibody immobilization line and a broad band of a complex of an anti-hCG-α antibody and gold colloid in a nitrocellulose film was manufactured as described below, and the liquid-impermeable sheet material was provided on the immuno-chromatography development layer and the base material was provided thereunder. Thereafter, an immuno-chromatography specimen which was cut employing a CO2 laser and an immuno-chromatography specimen which was cut with a cutting tool (score cutter) were manufactured to form an individual biosensor, and a measurement of hCG in urine was performed employing respective immuno-chromatography specimens to compare variation of their measurement values. [0144] A. Preparation of Immuno-Chromatography Development Layer [0145] First, the anti-hCG-β antibody solution which was diluted with a phosphate buffer solution to control the concentration was prepared. This antibody solution was applied on the nitrocellulose film by employing a solution discharge device. Thereby, a detecting antibody immobilization line was obtained on the nitrocellulose film. After being dried, this nitrocellulose film was immersed in a Tris-HCl buffer solution including a 1% skim milk, and gently shaken for 30 minutes. 30 minutes later, the film was moved into a Tris-HCl buffer solution tank, gently shaken for 10 minutes, and thereafter gently shaken in another Tris-HCl buffer solution tank for another 10 minutes, thereby washing the film. After washed twice as described above, the film was taken out from the cleaning fluid and dried at room temperature. [0146] The gold colloid was prepared by adding 1% citric acid solution to a refluxing 100� C.-solution of 0.01% chloroauric acid. After the reflux was continued for 30 minutes, the golod colloid was cooled and prepared to pH 9 by using a 0.2M potassium carbonate solution. The anti-hCG-α antibody was added to this gold colloid solution, then the obtained solution was stirred for several minutes, and thereafter a 10% BSA (bovine serum albumin) solution pH 9 was added thereto by such an amount that a 1% solution was finally obtained, and stirred. Thereby, an antibody-gold colloid complex (marked antibody) solution was prepared. Thereafter, this marked antibody solution was centrifuged at 4� C. and 20000G for 50 minutes, whereby the marked antibody was isolated, and the isolated marked antibody was suspended in a cleaning buffer solution (1% BSAphosphate buffer solution) and thereafter centrifuged to wash and isolate the marked antibody. After suspended in the cleaning buffer solution and filtrated through a 0.8 μm filter, the marked antibody was prepared to be one-tenth as much as the initial gold colloid solution and stored at 4� C. [0147] The so-created marked antibody solution was set in the solution discharge device and applied to a position apart from the antibody immobilization position on the anti-hCG-β antibody immobilization dry film, and thereafter the film was dried. Thereby, the marked antibody hold region was obtained on the immobilization film. [0148] In this way, the immuno-chromatography development layer can be completed. [0149] B. Creation of Immuno-Chromatography Specimen [0150] The immuno-chromatography development layer manufactured as described above was attached onto the base material made of white PET with 0.5 mm thickness, and the liquid-impermeable sheet made of transparent PET with 100 μm thickness was attached onto the development layer from the marked antibody holding part to the end of the development layer with a part for laminating the water absorbing part opened at the end. Then, there were manufactured the immuno-chromatography specimen which was cut with the width of 2.5 mm employing a CO2 laser, in which the side faces of the development layer are melted and cured to be sealed, and the immuno-chromatography specimen which was cut with a cutting tool (score cutter). Further, glass fiber filter paper is attached respectively to the ends as the water absorbing parts. In this way, two kinds of immuno-chromatography specimens can be manufactured. While in the example the glass fiber filter paper is attached as the water absorbing part, the water absorbing part is not indispensable, and it is also possible that the immuno-chromatography development layer except for its beginning, where the inspection target solution is applied, and end is adherently covered with the liquid-impermeable sheet, thereby providing the end of the development layer with the same effect as that of the water absorbing part. That is, by opening the end of the development layer, the inspection target solution is easily evaporated and it is utilized that the level of the inspection target solution which reached to the end of the development layer comes level with that of the inspection target solution in the beginning in relation to the water head difference. [0151] C. Preparation of Inspection Target Solution [0152] The hCG solutions of known concentrations were added to human urine, thereby preparing the hCG solutions of various known concentrations. [0153] D. Measuring Method of hCG in Urine [0154] More than 20 μl of urine including hCG was applied to sample application parts on the two kinds of immuno-chromatography specimens created as described above and developed in the direction of the water absorbing parts, to cause an antigen antibody reaction, whereby color reactions in antibody immobilization parts were caused. Here, the coloration states 5 minutes after the sample was applied to the specimens were measured by employing a reflective spectrophotometer (CS9300; Shimadzu Corporation made), and the coloration degree was computed. [0155]FIG. 10 are graphs illustrating relationships between hCG concentration and a measurement result in the above-described two kinds of immuno-chromatographic specimens according the example of the present invention; FIG. 10(a) illustrates the measurement result in the immuno-chromatographic specimen cut employing a CO2 laser and FIG. 10(b) illustrates the measurement result in the immuno-chromatographic specimen cut with a cutting tool (score cutter). [0156] First, urine including each hCG of hCG concentration 100, 1000, and 10000U/l were applied to the immuno-chromatography specimen to be developed. Then, the coloration state of the antibody immobilization part on the specimen for urine of each hCG concentration was measured by the reflective spectrophotometer. In the example, an absorbance at the wavelength of 520 nm was measured by the reflective spectrophotometer, and substituted into a previously formed calibration curve indicating a relationship between the hCG concentration and the absorbance, thereby obtaining a reduced value. [0157] As a result, CV values (coefficients of variation) of the specimen cut employing a CO2 laser shown in FIG. 10(a) were 3-10%, while the CV values of the specimen cut with a cutting tool (score cutter) shown in FIG. 10(b) varied widely between 20% and 35%. As described above, high quantitative accuracy was confirmed in the measurement employing the specimen cut employing a CO2 laser. APPLICABILITY IN INDUSTRY [0158] A biosensor and a manufacturing method thereof according to the present invention are quite useful as a biosensor which improves accuracy of a result of a reaction between a liquid sample as an analysis target and a marked reagent, and as a biosensor manufacturing method for simplifying a manufacturing process of such high-accuracy biosensor as well as reducing the cost. Referenced byCiting PatentFiling datePublication dateApplicantTitleUS7386937May 1, 2006Jun 17, 2008Roche Diagnostics Operations, Inc.Method of making a biosensorUS7476827Oct 29, 2004Jan 13, 2009Roche Diagnostics Operations, Inc.Method of making a biosensorUS7713750 *May 30, 2007May 11, 2010The Johns Hopkins UniversityAblation based laser machining of biomolecule patterns on substratesUS7780827Jun 22, 2005Aug 24, 2010Roche Diagnostics Operations, Inc.BiosensorUS8034609Nov 25, 2008Oct 11, 2011The Johns Hopkins Universitypatterning biomolecules on a substrate by coating the substrate with the biomolecules, applying a laser to the coating, and ablating a portion of the biomolecules with the laser in a predetermined pattern* Cited by examinerClassifications U.S. Classification435/7.9, 435/287.2, 156/251International ClassificationG01N30/90, G01N33/558, G01N33/543Cooperative ClassificationG01N33/558, G01N33/54393, G01N30/90European ClassificationG01N33/558, G01N33/543MLegal EventsDateCodeEventDescriptionNov 21, 2008ASAssignmentOwner name: PANASONIC CORPORATION, JAPANFree format text: CHANGE OF NAME;ASSIGNOR:MATSUSHITA ELECTRIC INDUSTRIAL CO., LTD.;REEL/FRAME:021897/0624Effective date: 20081001Owner name: PANASONIC CORPORATION,JAPANFree format text: CHANGE OF NAME;ASSIGNOR:MATSUSHITA ELECTRIC INDUSTRIAL CO., LTD.;US-ASSIGNMENT DATABASE UPDATED:20100203;REEL/FRAME:21897/624Free format text: CHANGE OF NAME;ASSIGNOR:MATSUSHITA ELECTRIC INDUSTRIAL CO., LTD.;US-ASSIGNMENT DATABASE UPDATED:20100209;REEL/FRAME:21897/624Free format text: CHANGE OF NAME;ASSIGNOR:MATSUSHITA ELECTRIC INDUSTRIAL CO., LTD.;US-ASSIGNMENT DATABASE UPDATED:20100216;REEL/FRAME:21897/624Free format text: CHANGE OF NAME;ASSIGNOR:MATSUSHITA ELECTRIC INDUSTRIAL CO., LTD.;US-ASSIGNMENT DATABASE UPDATED:20100309;REEL/FRAME:21897/624Free format text: CHANGE OF NAME;ASSIGNOR:MATSUSHITA ELECTRIC INDUSTRIAL CO., LTD.;US-ASSIGNMENT DATABASE UPDATED:20100316;REEL/FRAME:21897/624Free format text: CHANGE OF NAME;ASSIGNOR:MATSUSHITA ELECTRIC INDUSTRIAL CO., LTD.;US-ASSIGNMENT DATABASE UPDATED:20100323;REEL/FRAME:21897/624Free format text: CHANGE OF NAME;ASSIGNOR:MATSUSHITA ELECTRIC INDUSTRIAL CO., LTD.;US-ASSIGNMENT DATABASE UPDATED:20100330;REEL/FRAME:21897/624Free format text: CHANGE OF NAME;ASSIGNOR:MATSUSHITA ELECTRIC INDUSTRIAL CO., LTD.;US-ASSIGNMENT DATABASE UPDATED:20100406;REEL/FRAME:21897/624Free format text: CHANGE OF NAME;ASSIGNOR:MATSUSHITA ELECTRIC INDUSTRIAL CO., LTD.;US-ASSIGNMENT DATABASE UPDATED:20100413;REEL/FRAME:21897/624Free format text: CHANGE OF NAME;ASSIGNOR:MATSUSHITA ELECTRIC INDUSTRIAL CO., LTD.;US-ASSIGNMENT DATABASE UPDATED:20100420;REEL/FRAME:21897/624Free format text: CHANGE OF NAME;ASSIGNOR:MATSUSHITA ELECTRIC INDUSTRIAL CO., LTD.;US-ASSIGNMENT DATABASE UPDATED:20100427;REEL/FRAME:21897/624Free format text: CHANGE OF NAME;ASSIGNOR:MATSUSHITA ELECTRIC INDUSTRIAL CO., LTD.;US-ASSIGNMENT DATABASE UPDATED:20100504;REEL/FRAME:21897/624Free format text: CHANGE OF NAME;ASSIGNOR:MATSUSHITA ELECTRIC INDUSTRIAL CO., LTD.;US-ASSIGNMENT DATABASE UPDATED:20100511;REEL/FRAME:21897/624Free format text: CHANGE OF NAME;ASSIGNOR:MATSUSHITA ELECTRIC INDUSTRIAL CO., LTD.;US-ASSIGNMENT DATABASE UPDATED:20100525;REEL/FRAME:21897/624Free format text: CHANGE OF NAME;ASSIGNOR:MATSUSHITA ELECTRIC INDUSTRIAL CO., LTD.;REEL/FRAME:21897/624May 9, 2002ASAssignmentOwner name: MATSUSHITA ELECTRIC INDUSTRIAL CO., LTD., JAPANFree format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:NADAOKA, MASATAKA;TAKAHASHI, MIE;TANAKA, HIROTAKA;AND OTHERS;REEL/FRAME:012879/0429;SIGNING DATES FROM 20020219 TO 20020221May 1, 2002ASAssignmentOwner name: MATSUSHITA ELECTRIC INDUSTRIAL CO., LTD., JAPANFree format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:NADAOKA, MASATAKA;TAKAHASHI, MIE;TANAKA, HIROTAKA;AND OTHERS;REEL/FRAME:013407/0429;SIGNING DATES FROM 20020219 TO 20020221RotateOriginal ImageGoogle Home - Sitemap - USPTO Bulk Downloads - Privacy Policy - Terms of Service - About Google Patents - Send FeedbackData provided by IFI CLAIMS Patent Services