KR20000010701A - Chromatographic immunoassay device - Google Patents

Abstract

PURPOSE: A chromatography immunoassay device is provided, which can be used more easily, is protected from moisture and/or oxygen more effectively and can be produced at a lower cost than known chromatography immunoassay devices. CONSTITUTION: The chromatography immunoassay device is one in which one or more chromatography strips (1) are stuck on a substrate (2) made of a plate, each of the chromatography strips (1) is sealed by closely adhering a substrate portion surrounding each chromatography strip (1) to a seal film (3) located on the chromatography strip (1), and the seal film (3) and/or substrate possesses a film containing a dehumidifying agent and/or a film containing an oxygen absorbing agent, wherein chromatography strips (1) has an additional mark portion and the seal film (3) is a metal film.

Description

Immunochromatography Analyzer

Field of invention

The present invention relates to a chromatographic immunoassay device using a chromatography strip. More particularly, the present invention relates to a film comprising one or more chromatography strips positioned on a substrate, each chromatographic strip being sealed between the substrate and a sealing film covering the chromatography strip as a whole, the sealing film and / or the substrate containing a dehumidifying agent and / or A chromatographic immunoassay device having a film containing an oxygen absorber.

Background of the Invention

Chromatographic immunoassay devices using chromatographic strips are devices for immunologically detecting or measuring the presence or amount of a substance to be analyzed contained in a sample, the apparatus having at least sample addition means and detection means. In some cases, the chromatographic strips also have labeling means, such devices are described, for example, in document JP-A 61-145459, all of which are incorporated herein by reference (as used herein, the term “JP-A Means "Unexamined Japanese Patent Application", JP-A 1-63865 and JP-W 1-503174 (as used herein, the term "JP-W" refers to Unexamined Japanese Patent Application). International patent applications ", which are well known and widely used.

Each chromatography strip must be protected by appropriate means such as packaging to prevent degradation of antibodies and reagents, etc., in the chromatography strip by oxygen, moisture, etc. and to prevent the strip itself from being contaminated or modified by contact. Protection by packaging or similar means is generally accomplished by adhering or placing one or more chromatography strips onto a substrate, placing the resulting formulation in a protective case and then sealing the case in a bag. Alternatively, each chromatography strip is sealed with a laminated film as described in WO 94/24563, which is incorporated herein by reference. When packed in a bag, the chromatography strip is sealed with a dehumidifying agent to minimize moisture uptake by the chromatography strip.

However, in conventional chromatographic immunoassay devices, each of the chromatography strips is expensive to package because it requires a protective case, a dehumidifying agent and a wrapping paper to pack one chromatography strip. In the case of adhering multiple chromatography strips to a substrate, all chromatography strips are packaged in a single bag with the required amount of dehumidifying agent and sealed with a substantially waterproof packaging material. Although this method can reduce the cost required for packaging materials and dehumidifiers, once the bag is opened for use with the chromatography strip, there is a problem that other chromatography strips are exposed to air or moisture and oxygen and exacerbate them. Occurs. Therefore, special protection and storage measures are needed because the stored chromatographic strips become useless if not properly protected and stored. WO 94/24563, incorporated herein by reference, uses chromatographic strips that can be sequestered from air, but no further measures are provided to protect the chromatography strips from moisture and oxygen.

The gist of the invention

The present invention solves the above-mentioned problems in the prior art chromatographic immunoassay devices in terms of their form and packaging, making them easier to use and more effectively sequestered from moisture and / or oxygen for longer storage. The present invention provides a chromatographic immunoassay device capable of lowering the cost and lowering the cost.

Chromatographic immunoassays are being developed that are easier to use, more effective protection from moisture and / or oxygen, and have lower production costs. One or more chromatography strips are placed on a substrate, each chromatographic strip is sealed between the substrate and a sealing film covering the chromatography strip as a whole, the sealing film and / or the film containing a dehumidifying agent and / or an oxygen absorbent A chromatographic immunoassay device having a film containing

Accordingly, the present invention provides that (1) one or more chromatographic strips having at least sample addition means and detection means are located at regular intervals on a substrate consisting of one plate, with or without strip support, and (2) chromatographic strips. A sealing film covering the entirety, with or without a protective laminate, is placed in the aforementioned chromatographic strips, and (3) each chromatographic strip is brought into close contact with the substrate portion around each of the chromatographic strips. Sealing by adhesion, (4) the adhesion of the sealing film to the substrate mentioned above is carried out so that the sealing film can be easily peeled from the substrate at least in the sample addition region, and (5) as for the sealing film and the substrate, ) Both the sealing film or the substrate contains a film containing a dehumidifying agent and a film containing an oxygen absorbent (Ii) the sealing film has a film containing a dehumidifying agent or a film containing an oxygen absorbent, the substrate has another film, and (iii) the sealing film or substrate contains a film containing a dehumidifying agent or an oxygen absorbent A film, the other both having a film containing a dehumidifying agent and a film containing an oxygen absorbent, (iv) both the sealing film and the substrate each containing both a film containing a dehumidifying agent and a film containing an oxygen absorbent; (v) the sealing film and the substrate both contain the same film, which is a film containing a dehumidifying agent or a film containing an oxygen absorbent, or (vi) the sealing film or substrate contains a film containing a dehumidifying agent or a film containing an oxygen absorbent And (6) when the sealing film and / or the substrate has a film containing a dehumidifying agent, at least in a portion where the sealing film is not closely attached to the substrate. When the sealing film and the substrate are substantially waterproof, and the sealing film and / or the substrate have a film containing an oxygen absorbent, the sealing film and the substrate are substantially oxygen impermeable, at least in a portion where the sealing film is not closely attached to the substrate. The phosphorus chromatographic immunoassay device is a summary.

Effects of the Invention

It describes a chromatographic immunoassay device that is readily available, protects from moisture and / or oxygen, and has low production costs. The present invention provides a method of bonding one or more strips to a substrate made of a single plate, and sealing each chromatography strip by closely attaching a portion of the substrate surrounding each chromatography strip to a sealing film located on the aforementioned chromatography strip. The sealing film and / or the substrate is composed of a film containing a dehumidifying agent and / or a film containing an oxygen absorbent. Since chromatographic strips are isolated from moisture and / or oxygen by the sealing film and the substrate, they can be stored for a longer duration. Furthermore, even in the case of an apparatus in which a plurality of chromatography strips are adhered to a substrate, when using one chromatography strip, the remaining chromatography strips are not exposed to air, unlike the conventional type of apparatus currently used. The chromatographic strips in the present invention can also be separated from other strips together with the substrate without exposure to air before or after use. In addition, the chromatographic immunoassay device of the present invention does not require the labor of packaging required for chromatographic strip preparation in the prior art. Thus, the strips described herein can be produced at less cost.

Detailed description of the invention

In the chromatography strip, at least sample addition means and detection means are arranged on the chromatography carrier. The sample solution, which may contain the substance to be analyzed, moves through the chromatographic carrier by capillary action when applied to the sample addition means, and the labeling material or sample solution contained in the label means previously arranged in the chromatography strip. Since the labeling substances added to the chromatography strip together accumulate in the detection means in direct proportion to or inversely proportional to the presence or amount of the substance being analyzed in the sample solution resulting from the immunological reaction, the presence or amount of the substance being analyzed in the sample solution is accumulated. By measuring the presence or amount of labeling substance. Various types of chromatography strips are known and all such known chromatography strips can be used in the present invention, including those described later. As used herein, the term "chromatographic immunoassay device" refers to a chromatography strip that can be used for immunoassay and prepared for storage and transport.

Next, an example of a conventional chromatography strip is described. If a labeling means is present, the sample addition means may be located at the same place where the labeling means are present or at an upstream position relative to the labeling means (hereinafter the direction of movement of the sample solution generated by capillary action is "Downstream" and the opposite direction is referred to as "upstream." In general, upstream to the labeling means is preferred. When introducing a sample solution that may contain the substance to be analyzed into the sample addition means, the sample solution moves with the substance to be analyzed through the chromatography carrier in the downstream direction generated by capillary action. Typically, the substance to be analyzed is a compound that binds in a specific way to a trapping substance fixed to the detection means, or a compound that binds in a specific way to a conjugate that specifically binds to the trapping material. For example, the substance to be analyzed is an antibody if the capture substance is an antigen or the conjugate contains an antigen, and the substance to be analyzed is an antigen if the capture substance is an antibody or the conjugate contains an antibody.

When the sample adding means is located upstream with respect to the labeling means (when a labeling means is present), the labeling means may be arranged adjacent to or apart from the sample adding means. Labeling materials that specifically bind to the material to be analyzed or that specifically bind to the capture material in competition with the material to be analyzed are arranged on the labeling means.

If there is no labeling means, the labeling substance can be added to the sample addition means together with the sample solution, but the addition of the labeling substance, for example, after adding the sample solution, places the labeling material at a specific location outside the chromatographic strip binding site. By addition, it can carry out by various methods.

The label may be a coloring substance such as a radioisotope, enzyme or gold colloid or the like. Such labels are also well known.

Since the labeling material is arranged in a manner that moves by capillary action of the sample solution, when the sample solution is added to the sample adding means, the labeling material moves in the downstream direction.

In general, the detection means is located downstream from the labeling means and at a distance from the labeling means. In the detection means, the capture material which binds only to the substance or conjugate to be analyzed in a specific manner or which specifically binds to the substance to be analyzed and the labeling substance, respectively, is immobilized on the chromatography carrier. As a result, in one embodiment, the material to be analyzed (sometimes linked to the labeling material), which is moved by capillary action of the sample solution, binds to the capture material or conjugate and then to the capture material. Thus, since the labeling substance binds to the substance to be analyzed, the labeling substance is accumulated in the detection means in response to the presence or amount of the substance being analyzed. In contrast, the substance to be analyzed with the label material, which is moved by capillary action, binds to the capture material or conjugate and then competitively binds to the capture material, causing the label material to accumulate in inverse proportion to the amount of material being analyzed.

Any labeling substance, if not simultaneously, may bind to both the capture substance (or to the conjugate and then to the capture substance) and the substance to be analyzed, in which case the substance to be analyzed first binds to the labeling substance. And the remaining labeled material that does not bind to the material being analyzed binds to the capture material. As a result, the presence or amount of the substance to be analyzed can be analyzed by measuring the label substance accumulated in the detection means.

In some cases, several substances are located upstream of the detection means. For example, the conjugate may be located in a movable manner. A conjugate is a complex of a compound that specifically binds to a substance or labeling substance to be analyzed and another compound that specifically binds to a capture substance, which is a labeling substance and a capture substance to both the substance to be analyzed and the capture substance or in a specific manner. To combine. Examples of combinations of capture materials and compounds that specifically bind to the corresponding capture materials include biotin and avidin (both may be capture materials), antibodies and their corresponding antigens (both not related to the sample being analyzed). Etc. are included.

In some cases, one or more additional detection means may be arranged downstream of the first detection means. In addition, the downstream of the detection means here may further expand the chromatography carrier to allow complete drainage of the sample solution or the carrier to be composed of the material used to absorb the sample solution.

Chromatographic carriers are carriers of the sample addition means, the labeling means and the detection means, which connect these means so that the sample solution can be moved by capillary action. Many materials have been proposed as chromatography carriers, and any of these materials can be used as the chromatography carrier of the present invention. For example, cellulose, nitrocellulose, cellulose acetate and the like are most frequently used as chromatography carriers.

Thus, the presence or amount of the substance to be analyzed in the sample solution can be known by measuring the presence or amount of labeling substance accumulated in the detection means. In one example, this can be done visually.

If desired, the chromatography strip may be attached to the strip support such that one side thereof is in contact with the strip support (hereinafter, the side that contacts the strip support is referred to as the "back side" of the chromatography strip). The strip support is mainly used to prevent the sample adding means, the labeling means and the like from moving. Adhering the chromatography strip to the strip support should be carried out so that the capillary action of the sample solution in the chromatography carrier is not disturbed so that the detection sensitivity of the material being analyzed is not reduced. In some cases, the strip support is used so as not to cover the sample addition means portion.

Also, if desired, the protective stack may adhere to the opposite side of the strip support-adhesion side of the chromatography strip [hereinafter, the face, or the opposite side to the substrate-adhesion side, may be the “front side” of the chromatography strip. Called. Protective laminates are mainly used to ensure the attachment of sample addition means and labeling means and to prevent stains and other gold from occurring in the chromatographic strip when used. At least one portion of the protective stack should be transparent when covering the detection means, and the sample addition portion of the sample addition means should not be covered by the protective stack. The adhesion of the chromatography strip to the protective stack should be carried out so that the capillary action of the sample solution on the chromatography carrier is not disturbed so that the detection sensitivity of the material being analyzed is not reduced.

Polyethylene terephthalate (hereinafter referred to as "PET") is most frequently used as strip support and protective laminate. Polypropylene (hereinafter referred to as "PP"), polyvinyl chloride and the like may also be used.

The attachment of the chromatographic strip to the strip support or protective laminate or to the substrate described later is carried out by using a rubber, acrylic or vinyl ether polymer adhesive.

One or more chromatographic strips may be placed on the strip substrate with or without strip support. As used herein, the term “locating” simply places the chromatography strips on the substrate, or in other cases, adheres them to the strip support, if present, or to the substrate if there is no strip support. It means The term "adhesive" as defined herein means to attach the entire surface or only a portion of the chromatography strip to the substrate. In any case, the adhesion of the chromatographic strip is effective when it is not easily separated from the strip support or substrate during or during its use. In some cases, the substrate may be adhered with a paste so that it can be easily peeled off from the chromatography or strip support.

If no strip support is used, the substrate may have the function of a strip support.

When multiple chromatography strips are placed on a substrate, in terms of production, it is desirable to equalize the downstream and upstrip ends of each chromatography strip so that they are parallel. Such chromatographic strips are placed on the substrate and spaced apart.

The substrate, if present, consists of a single plate and adheres to the strip support or to the back side of the chromatography strip.

The chromatographic strip seals the sealing film described later by closely adhering to the substrate at the parallel plane of each chromatography strip positioned on the substrate, i.e. in the space between the chromatographic strips when multiple strips are located on the substrate. .

The sealing film is a sheet of film that can cover the entire portion of the chromatography strip and is placed on the chromatography strip with or without a protective laminate.

When the substrate is closely adhered to the sealing film by hot sealing, the inner surface of the substrate (the surface where the chromatography strip exists) and the inner surface of the sealing film (the side where the chromatography strip exists) must be hot sealable. That is, they must contain a material that can be hot sealed. Examples of such sealable films and hot sealable materials for substrates include a combination of a film containing polyethylene (hereinafter referred to as "PE") or PP on its inner surface with a film whose inner surface is coated with a corresponding hot melt adhesive; or A combination of a film containing PE on the inner surface and a film containing a PP-PE copolymer film on the inner surface.

When the sealing film is adhered to the substrate as a paste, this may be carried out by a combination of any sealing film and a substrate containing a rubber, acrylic acid or vinyl ether polymer adhesive on the inner surface.

The close adhesion of the sealing film and the substrate should be made so that the substrate and the sealing film can be easily peeled off when used, at least in the sample addition means portion. In order to easily peel off the substrate and the sealing film and maintain proper sealing performance, peel strength of 1.5 to 2.0 kg per 15 mm width is preferred.

As for the sealing film and the substrate, (i) the sealing film or the substrate has both a film containing a dehumidifying agent and a film containing an oxygen absorbent, and (ii) the sealing film contains a film containing a dehumidifying agent or an oxygen absorbent. A film having a film, the substrate having another film, and (iii) the sealing film or the substrate having a film containing a dehumidifying agent or a film containing an oxygen absorbent, the other being a film containing a dehumidifying agent and a film containing an oxygen absorbent. Both of which have both (iv) a film containing a dehumidifying agent and a film containing an oxygen absorbent, and (v) a film or an oxygen absorbent both containing a dehumidifying agent (Vi) The sealing film or the substrate contains a film containing a dehumidifying agent or a film containing an oxygen absorbent. When the sealing film and / or the substrate has a film containing a dehumidifying agent, the sealing film and the substrate are substantially waterproof, at least in a portion where the sealing film does not adhere closely to the substrate, and the sealing film and / or the substrate contains an oxygen absorbent. In the case of having a film, the sealing film and the substrate are substantially oxygen impermeable, at least in a portion where the sealing film is not closely attached to the substrate.

When the sealing film has a film containing a dehumidifying agent, the sealing film has a waterproof layer on the outside. In addition, when the sealing film has a film containing an oxygen absorbent, the sealing film has an oxygen impermeable layer on the outside. In the same way, when the substrate has a film containing a dehumidifying agent, the substrate has a waterproof layer on the outside, and when the substrate has a film containing an oxygen absorbent, the substrate has an oxygen impermeable layer on the outside. In many cases, the outermost surface of the sealing film is made ready for printing thereon.

If the sealing film and / or substrate does not have a film containing a dehumidifying agent and a film containing an oxygen absorbent, the sealing film and / or the substrate is an oxygen impermeable film, a waterproof film, a film suitable for hot sealing, the outermost PET It may be a single layer film or a multilayer film obtained by any combination of films and the like.

If both the substrate and the sealing film do not have an oxygen absorbent containing film, the substrate and the sealing film need not be oxygen impermeable, and if both the substrate and the sealing film do not have a dehumidifying agent containing film, the substrate and the sealing film need to be waterproof. There is no. That is, when the chromatography strip is not decomposed by moisture, there is no need to use a dehumidifying agent-containing film or a waterproof sealing film, or when the chromatography strip is not decomposed by oxygen, an oxygen absorber-containing film or an oxygen permeable sealing film and There is no need to use a substrate.

Dehumidifier-containing films are thermoplastic high molecular weight resins, preferably polyolefins, more preferably low density polyethylene, linear low density polyethylene, ethylene-vinyl oxide copolymers, ethylene acrylic acid copolymers, ethylene methacrylic acid copolymers, ethylene acrylic ester copolymers and One selected from acrylic and methacrylic acid copolymer-based ionomers can be prepared by kneading an appropriate amount of calcium chloride, silica gel, molecular sieve, silicon dioxide, alumina, zeolite magnesium sulfate, gypsum and the like used as a drying agent. Examples of dehumidifying agent-containing films include "Moisture Guard" (Toyo Seikan) and "Highsheet Dry" described in Japanese Patent Publication No. 08-026348 (26348/96), which is incorporated herein by reference. Film "(Hiseat-Dry Film, manufactured by Marutani Kakoki). A particularly preferred dehumidifier containing film is 110 μm, and a desiccant layer of about 90 μm containing low density PE (LDPE) of about 10 μm, various amounts of positive high molecular weight resins (eg LDPE) and desiccants such as zeolites, molecular sieves And LDPE of about 10 μm. Preferably the suitable dehumidifier containing film contains a layer having a desiccant content of about 0.1 to 50% by weight, preferably 10 to 50% by weight. Particular preference is given to a total desiccant content of about 8 to 50 g / m ² . It is desirable to prepare a layer containing a desiccant using a desiccant having an average particle size of about 5 to 70 microns.

The oxygen absorbent-containing film can be produced by kneading a thermoplastic high molecular weight resin with an appropriate amount of activated iron oxide, pyrogallol, an oxygen absorbent and the like. An example of an oxygen absorber containing film is "Oxy Guard" by Toyo Seiken.

In some cases, the sealing film is a film having both a dehumidifying agent and an oxygen absorbent. Sealing films of this type may be referred to herein as dehumidifier containing films or oxygen absorbent containing films. In the same way, the substrate may be a film having both a dehumidifying agent and an oxygen absorbent. Substrates of this type may be referred to herein as dehumidifier containing films or oxygen absorbent containing films.

An example of a substantially waterproof substrate is a multilayer consisting of at least 300 μm PE, at least 300 μm PP, at least 300 μm [PE or PP] from the inner surface and about 15 μm polyvinylidene chloride (hereinafter referred to as “PVDC”). Film, a multilayer film consisting of about 70 μm [PE or PP] from the inner surface, about 125 μm PET and about 15 μm PVDC. Examples of substrates that are substantially waterproof and contain a dehumidifying agent containing film include a “moist guard” of 300 μm and a “moist guard” of about 110 μm from the inner surface, a multilayer film consisting of about 125 μm PET and about 15 μm PVDC. have. An example of a substrate that is substantially oxygen impermeable is a multilayer film consisting of at least about 150 [mu] m of [PE or PP], about 15 [mu] m of polyvinyl alcohol saponification and at least 150 [mu] m of [PE or PP] from the inner surface. The above examples are all transparent.

An example of a substantially waterproof and oxygen impermeable substrate is a multilayer film consisting of at least 150 μm [PE or PP], about 30 μm PVDC and at least 50 μm [PE or PP] from the inner surface. An example of a substantially waterproof, oxygen-impermeable and opaque substrate is a multilayer consisting of at least 200 μm [PE or PP] from the inner surface, an aluminum foil of about 7 μm (hereinafter referred to as “Al”) and a PET of about 15 μm Film, 70 μm [PE or PP] from the inner surface, about 125 μm PET, about 7 μm Al and 12 μm PET, and about 70 μm [PE or PP] from the inner surface, about 125 There is a multilayer film consisting of polystyrene of μm, Al of about 7 μm and PET of about 12 μm.

Preferred substrates consist of a layer of about 30 μm which facilitates the removal of the sealing film from the inner surface, a blend of PE and PP, about 188 μm white PET (for color contrast), and about 7 μm of Al (oxygen and moisture). Barrier) and about 12 μm PET. Particularly preferred substrates consist of a layer of about 30 μm which facilitates the removal of the sealing film from the inner surface and consists of a blend of PE and PP, about 50 μm white PET, about 7 μm Al and about 188 μm PET. .

Substrates that are substantially waterproof and oxygen impermeable and that contain a dehumidifying agent and / or oxygen absorbent containing film are substantially waterproof and oxygen impermeable to the " moisture guard " It can be obtained by replacing with "oxy guard" of 110-250 μm.

If desired, a rubber, acrylic or vinyl ether polymer system may be applied to the sides of the substrate, in particular the inner side. In this case, it is laminated on the paste-coated side until the release paper or release film is used. It can be used as a release paper or a release film without particularly limiting paper, PET or PP.

An example of a substantially waterproof sealing film is a multilayer film consisting of [PE or PP] of about 70 μm and PET of about 12 μm from the inner surface. Substantially oxygen impermeable is a multilayer film consisting of about 70 μm [PE or PP], about 15 μm polyvinyl alcohol saponifier, about 12 μm PP, and about 12 μm PET from the inner surface. Substantially waterproof and oxygen impermeable is a multilayer film consisting of about 70 μm [PE or PP], about 30 μm PVDC and about 12 μm PET from the inner surface. The above examples are all transparent.

An example of a sealing film that is substantially waterproof and contains a dehumidifier containing film is a multilayer film (transparent) consisting of a “moisture guard” of 110 μm from the inner side and a PET of about 12 μm, particularly preferably of about 110 μm from the inner side. "Moisture Guard", a multilayer film (opaque) consisting of Al of about 7 μm and PET of about 12 μm. An example of a sealing film that is substantially oxygen-impermeable and contains an oxygen absorber-containing film is a multilayer film (opaque) consisting of 110 micrometers of "oxyguard", 7 micrometers of Al and 12 micrometers of PET from the inner surface. Examples of sealing films that are substantially waterproof and oxygen impermeable and contain a dehumidifier containing film and an oxygen absorbent containing film include a “moisture guard” of 110 μm, an “oxy guard” of 110 μm, Al and 12 μm from an inner surface. It is a multilayer film (opaque) made of PET of μm.

Multilayer films can be prepared by adhering the constituent film with an appropriate adhesive or coextrusion of a laminated portion of the constituent film, and then optionally bonding the residual constituent material with an appropriate adhesive.

The chromatographic immunoassay device first prepares the above-mentioned chromatography strips and, if appropriate, is prepared using the strip support and / or protective laminate in the above-mentioned manner, or to prepare the chromatography strips and at the same time sample addition means. Or the like, and the above-mentioned substrate is firmly attached to the sealing film to seal the chromatography strip.

The chromatographic immunoassay device of the present invention is used in the following manner. Only one chromatographic strip used is cut with the substrate. One or more portions covering the sealing film or the sample addition means are peeled off and the sample solution is added to the exposed sample addition means. In the case of certain shapes of the sealing film, it may be necessary to peel off the entire portion of the sealing film covering the chromatography strip. When multiple chromatography strips are placed on a substrate and used without first cutting the chromatography strips, they are generally cut after use. In some cases, the device can be used by separating the substrate from the strip support.

The invention is further described with reference to the drawings. 1A, 1B and 1C are schematic diagrams showing examples of the chromatographic immunoassay apparatus of the present invention. 1A is a plan view, FIG. 1B is a sectional view, and FIG. 1C is another sectional view. 2 is a schematic diagram illustrating an example of a chromatography strip. 3 is a schematic view showing still another example of the chromatographic immunoassay device of the present invention. In these figures, (1) represents a chromatography strip, (2) represents a substrate, (3) represents a sealing film, (4) represents a chromatography carrier, and (5) represents a sample addition means. (6) represents labeling means, (7) represents detection means, (8) represents strip supports, (9) represents protective laminates, (10) represents perforations, and (11) ( Diagonal lines indicate the closely attached portions of the substrate and the sealing film.

The chromatography strip (1) is located on the substrate (2) and has a film containing a dehumidifying agent and / or an oxygen absorbent, and the chromatography strip (1) has a sealing film (3) on the periphery of the chromatography strip of the substrate (2). 11) to seal and isolate from air (13).

When using a comb-shaped substrate as shown in FIG. 3, the sample addition means of each chromatography strip are completely separated, eliminating the possibility of sample solution accidentally flowing into adjacent chromatography strips when adding the sample.

The chromatography strip 1 is constituted by arranging the sample addition means 5, the labeling means 6 and the detection means 7 on the chromatography carrier 4. If desired, the chromatography strip is supported or protected by the strip support 8 and the protective stack 9.

The perforations 10 are optionally arranged when necessary to easily cut one chromatography strip from the chromatography immunoassay device when multiple chromatography strips are located on the substrate. Perforation 10 may be used to cut one chromatography strip or to cut several chromatography strips simultaneously.

1A, 1B and 1C are schematic diagrams showing examples of the chromatographic immunoassay apparatus of the present invention, wherein FIG. 1A is a plan view, FIG. 1B is a side elevation view, FIG. 1C is an elevation view,

2 is a schematic diagram showing an example of a chromatography strip,

3 is a schematic view showing still another example of the chromatographic immunoassay device of the present invention.

Code Description

(1) chromatography strip

(2) substrate

(3) sealing film

(4) chromatography carriers

(5) sample addition means

(6) marker means

(7) detection means

(8) strip support

(9) protective laminate

10 perforation

(11) closely attached portions of the substrate and the sealing film

Example 1-Performance of Seal Film and Substrate Combination

The combination of the sealing film and board | substrate shown in Table 1 is manufactured. Multilayer films are each produced by adhering individual monolayer films together with an adhesive.

Sample number Sealing film Board One MG / Al / PET ₁₂ PP ₇₀ / PET ₁₂₅ / Al / PET ₁₂ 2 MG / PVDC / PET ₁₂ PP ₃₀₀ 3 MG / Al / PET ₁₂ MG / PET ₁₂₅ / Al / PET ₁₂ 4 MG / OG / Al / PET ₁₂ PP ₇₀ / PET ₁₂₅ / Al / PET ₁₂ 5 OG / Al / PET ₁₂ MG / PET ₁₂₅ / Al / PET ₁₂ 6 MG PP ₃₀₀ 7 (control) PP ₇₀ / Al / PET ₁₂ PP ₇₀ / PET ₁₂₅ / Al / PET ₁₂

MG: Moisture Guard 110μm (Toyo Seikan)

OG: "oxy-guard" of 110 μm (Toyo Seikan)

Al: 7 μm Al

PET ₁₂ : 12㎛ PET

PET ₁₂₅ : 125 μm PET

PP ₇₀ : 70㎛ PP

PP ₃₀₀ : 300㎛ PP

PVDC: 12µm PVDC

A. Dehumidification Performance Test 1

Sealing films of samples 1, 2, 3, 4 and 7, substrates of samples 3 and 5 and commercially packaged granulated silica gel in individual amounts shown in Table 2 were subjected to constant temperature of room temperature (23 ° C., 40% humidity) or 40 ° C. Store in oven for 3 days. The weight of each sample is then measured and the amount increased from the initial weight is used as the water uptake. The results are shown in Table 2.

These results demonstrate that films with dehumidifier containing films have a high dehumidification capacity.

Sample Sample amount Water absorbed (mg) 23 ℃ 40 ℃ Sample 1 sealing film 300 cm ² 84.2 79.7 Seal Film of Sample 2 300 cm ² 84.0 79.5 Sample 3 sealing film 300 cm ² 83.5 78.9 Sample 4 sealing film 300 cm ² 83.7 78.6 Substrate of Sample 3 300 cm ² 84.9 80.3 Substrate of Sample 5 300 cm ² 84.1 79.9 Packaged Granular Silica Gel (Control) 5 g 892 288 Seal Film (Control) of Sample 7 30 cm ² 0.1 -0.1

The above study is repeated comparing the absorption capacity of silica gel (5 g package) with the sealing film of Sample 1 or 3 containing 0.75 g of desiccant / 300 cm ² (sealed film A) or 1.50 g. Storage conditions are 6 days at 24 ° C. and 50% relative humidity, 40 ° C. and 20% relative humidity or 2 to 8 ° C. and 90% relative humidity. The results (Table 3) are calculated as above (mg of water absorbed) and also as absorbed water (g) / desiccant (g%). As can be seen from these results, the film has a high moisture absorption, dehumidification capacity. Also, film B, which contains twice as much desiccant than film A, can absorb approximately twice as much water as film A.

Sample Storage condition (temperature / relative humidity) Save time (days) Absorbed water (mg) (g / g%) Sealing Film A Sealing Film A Sealing Film A Sealing Film A 40 ° C./20% 24 ° C./50% 2 to 8 ° C./90% 2 to 8 ° C./90% 66619 12914176144 17.2618.7810.1719.13 Sealing Film B Sealing Film B Sealing Film B Sealing Film B 40 ° C./20% 24 ° C./50% 2 to 8 ° C./90% 2 to 8 ° C./90% 66619 268292121277 17.8419.458.0618.47 Silica gel silica gel silica gel silica gel 40 ° C./20% 24 ° C./50% 2 to 8 ° C./90% 2 to 8 ° C./90% 66619 495147218571937 9.929.4437.1538.74

B. Dehumidification Performance Test 2

Using the sealing films of Samples 1, 2, 3, 4 and 6 and the substrates of Samples 3 and 5 (see Table 1), a bag was prepared (inner surface area 600 cm ² ) and the three sides of the bag were hot sealed. Each bag of humidity indicator paper manufactured by Humidal Company, USA is placed in each bag.

Then heat seal the remaining side. A similar bag (inner surface area 600 cm ² ) was made using the sealing film of Sample 7. 2 g of the granular silica gel packed with the humidity indicator paper described above are placed in a bag and the remaining side is hot sealed. As a control, a similar bag (inner surface area 600 cm ² ) is prepared using the sealing film of Sample 7, the humidity indicator paper described above is placed in the bag and the remaining side is sealed by hot sealing. The prepared bag is then left to stand at room temperature for 3 days and then opened and immediately read the humidity indicator paper as soon as the bag is opened.

The humidity indication is 45% in the control bag and less than 15% in all other bags. The minimum humidity measurable with the humidity indicator paper is 15%,

C. Oxygen Absorption Performance Test

Bags were prepared using the sealing films of Samples 4 and 5 (see Table 1) (inner surface area 600 cm ² ) and the four sides of each bag were hot sealed. A piece of natural rubber 1 x 1 cm is attached to the surface of the bag with an adhesive. Using a syringe, the air is completely removed from the bag through a piece of natural rubber and then 20 ml of air is injected correctly. After 3 days at room temperature, the oxygen concentration is measured using gas chromatography, which removes the air portion of the bag through a piece of natural rubber and packs the column with molecular sieves.

No oxygen was detected in any of the bags tested. The oxygen concentration detection limit in this analysis system is 0.01%. These results demonstrate that a multilayer film with an oxygen absorber containing film has an oxygen absorption capacity.

Based on the above results, a chromatographic immunoassay device prepared by the method described in Example 2 below using a sealing film and a substrate of Samples 1, 2, 3, 4, 5 or 6, has a high dehumidification capacity. Will have Chromatographic immunoassay devices prepared using the sealing films and substrates of Samples 4 or 5 are also expected to have high oxygen uptake capacity.

Example 2 Chromatographic Immunoassay Device Stability

A. Device Manufacturing

The labeling substance consisting of anti-human hemoglobin antibody labeled with selenium colloid is prepared by the following method. First, selenium colloid is prepared by stirring 91 mM sodium L-ascormate and 32 mM selenium oxide at about 4 ° C. for 15 minutes and then at about 42 ° C. for about 70 hours. The selenium colloid obtained is thus diluted with 10 mM bis-tris buffer (pH 7.0) to give an absorbance of 15 at 550 nm. The resulting dilution is mixed with mouse monoclonal antihuman hemoglobin antibody (0.02%) and stirred at room temperature for 1 hour. The selenium colloid-labeled anti human hemoglobin antibody thus obtained is washed with 10 mM Tris-HCl buffer (pH 7.2) and used as labeling material.

Labeling means are prepared by the following method: Labeling material is added to 10 mM Tris-HCl buffer (pH 7.2) containing 1% casein and the absorbance at 550 nm is adjusted to 1.0 to obtain a labeling substance suspension. A glass fiber membrane (trade name: Lypore 9524, manufactured by LYDALL, U.S.A.) is immersed in the obtained suspension and sufficiently impregnated, and then the glass fiber membrane is dried to use as a labeling means.

The detection means are prepared by the following method: 30 mM Tris- containing 150 mM of sodium chloride at a final concentration of 2 mg / ml of a mouse monoclonal antihuman hemoglobin antibody whose binding site to human hemoglobin differs from the binding site of the antihuman hemoglobin antibody mentioned above. Add to HCl buffer (pH 7.4). Separately, a 0.4 x 4.5 cm rectangular piece of nitrocellulose membrane (Scheleicher & Schuell, USA) with a pore size of 5 μm as a chromatography carrier was prepared using a strip support (PE PET100 PE with a rectangular size of 0.4 × 6.0 cm and a thickness of 100 μm). LR007A, Lintech, Inc., so that when the longer side is arranged vertically, the two pieces of downstream ends match each other. The solution mixed with the antibody is added dropwise to the nitrocellulose membrane adhered to the support to form a string about 1 cm from the upstream end of the membrane. It is dried sufficiently to allow the anti-human hemoglobin antibody to be immobilized on nitrocellulose.

Chromatography strips are prepared by the following method. The labeling means mentioned above is cut into 0.4 x 0.4 cm square pieces and glued upstream of the strip support of the detection means containing the anti-human hemoglobin fixed nitrocellulose membrane to slightly contact the nitrocellulose membrane. A nonwoven fabric (Sontara 8801, Du Pont) cut to 0.4 x 1.3 cm as a sample addition means is adhered to the strip support upstream of the label means so as to be slightly in contact with the label means. A protective film having a rectangular size of 0.4 x 5.1 cm is adhered on this, and when the longer side is arranged vertically, the upper end is matched with the nitrocellulose membrane to obtain a chromatographic strip.

The chromatography strips are adhered to the substrate in the following manner: substrates of samples 1 or 7 (see Table 1) in parallel at 1.8 cm intervals by uniformly arranging the upper ends of the strip supports of each of the ten chromatography strips To

The sealing film is closely adhered to the substrate by hot sealing in the following manner: The substrate to which the chromatography strip is bonded is divided into two equal parts, and the 0.25 cm perimeter around each chromatography strip is not hot sealed. The part is hot sealed with the sealing film of Example 1. Hot sealing is carried out under a sealing temperature of 120 ° C., a sealing period of 1.5 seconds and a sealing pressure of 3.0 kg / cm ² . It is further divided into two parts and one of them is punched as shown in FIG. 1 using a cutting die. Use this as device A. Also using a cutting device, the remainder is made in device B with one chromatography strip.

As a control, the remaining portion of the substrate to which five chromatographic strips were bonded together with 5 g of commercially available packaged granular silica gel was placed in a bag (25 × 15 cm size) obtained by hot sealing three sides of the sealing film of Sample 7 The bag is hot sealed. This is used as the device C.

As another control, the substrate of Sample 6 to which the chromatography strips are adhered is hot sealed with the sealing film of Sample 7 in the same manner as described above and used as Apparatus D.

B. Store under extreme conditions

The prepared devices A, B, C and D are left for 1 day at 25 ° C. under 60% relative humidity and then for 28 days at 40 ° C. under 70% relative humidity. After storing the devices A, B, C and D under these extreme conditions, they were left for 2 hours at 25 ° C. and then opened while the devices A, B and D were left unchanged and the device C exposed to the outside air. Leave at 25 ° C. for 24, 48 and 96 hours at 60% wet.

Sample solutions were human hemoglobin (Sigma, USA) 0, 5, 10, 25, 50, 100, 200 or 500 ng / ml, 0.1% bovine serum albumin (Seikagaku Kogyo), 0.9% sodium chloride and 0.1M Tris- Prepared by dissolving in 0.1% sodium azide in HCl buffer (pH 7.6). 25 μl of the prepared sample solutions are added to the chromatographic strip sample addition means of each of the devices A, B, C and D 24, 48 or 96 hours before, immediately after or after storing them under extreme conditions. The result is judged by observing the serene colloid-origin "yellowing" which appears when the sample is detected positively by the detection means of the strip 7 minutes after adding the sample solution. Sensitivity is based on the minimum hemoglobin concentration at which "yellowing" is observed by the naked eye. The results are shown in Table 4.

Device Before and after storage under extreme conditions I'm Immediately after 24 hours later 48 hours later 96 hours later A 25ng / ml 25ng / ml 25ng / ml 25ng / ml 25ng / ml B 25ng / ml 25ng / ml 25ng / ml 25ng / ml 25ng / ml C 25ng / ml 25ng / ml 25ng / ml 50ng / ml 100ng / ml D 25ng / ml 200ng / ml 200ng / ml 500ng / ml 500ng / ml

As is apparent from Table 4, immediately after storage under extreme conditions the chromatographic strips of devices A, B and C exhibit the same sensitivity as the chromatography strips before storage under extreme conditions, and their stability during storage under extreme conditions is excellent. Do. As the storage period under extreme conditions proceeds to 24 hours, 48 hours and then 96 hours, the sensitivity of devices A and B does not change but the sensitivity of device C gradually decreases. This indicates that the unsealed device C is poor in stability. Apparatus D, which contains no dehumidifier-containing film or desiccant at all, shows that its sensitivity is significantly reduced by extreme condition tests.

As can be seen from this test, conventional products are required to be stored in a cold room since some chromatographic strips cannot be stored at room temperature. A device manufactured by a conventional method cannot be operated quickly if, for example, an urgent test is required because it is necessary to first warm the device to room temperature in order to perform analysis after storing it in a cold room. However, since device A can be stored at room temperature, it can cope with an emergency situation. In addition, the stability of the chromatography strip can be used with confidence since it can be ensured until the sealing film is peeled off. Since the sealing film can be easily peeled off, no hard labor is required when using the device A.

In terms of production costs, prior art products need to package and seal a device manufactured with a dehumidifying agent or the like in a substantially waterproof bag, while device A does not require such operation, so the production cost is greatly reduced and slightly This can offset the increase in substrate and sealing film costs. The manpower and associated costs required to manufacture device A may also be lower than current products.

C. 1 month stability

Chromatography strips for detecting human hemoglobin were prepared as in Example 2.A above and 13 strips were made free of desiccant (A), 110 μm of MG [10 μm of PE / PS (PS is described herein below). Defined as polystyrene), made of a 90 μm desiccant layer containing 11.9 g / m ² and 10 μm PE / PS] / packed with 15 μm Al / 12 μM PET (B), the same seal of 678.6 cm ² It is packaged in an aluminum bag packed with a film (C) or an aluminum bag having a conventional desiccant consisting of 1.3 g of silica gel (D). All strips are exposed to 65% relative humidity overnight at 25 ° C. prior to packaging and heat sealing. The silica gel and sealing film used are exposed to 65% relative humidity at 25 ° C. for 6 hours before use.

After heat-sealing the strips under the packaging conditions described above, they are stored in an incubator at 25 ° C. and tested after 16 and 29 days (25 C storage) or stored in a 25 ° C. incubator for 1 day and then transferred to an incubator at 37 ° C. and 37 ° C. Test 15 and 28 days after transfer to (store 37C).

The test is carried out using samples containing human hemoglobin 0 (negative control), 10, 25, 50, 100 or 500 ng / ml as in B. The results are shown in Table 5 as the minimum hemoglobin concentration at which 7 minutes after the sample was added and the signal can be visually observed.

Packing condition Test sensitivity (ng / ml): 25C storage 37C storage 16th 29 days 16th 29 days A (no desiccant) 25 50 100 500 B (1 × Sealed Film) 25 25 25 25 C (10 × Sealed Film) 25 25 25 25 D (silica gel) 25 25 25 25

Strips that do not contain any desiccant show poor sensitivity and thus show poor stability after storage at 25 ° C. or 37 ° C. for 1 month. When the strip is packaged with a desiccant, no degradation in test sensitivity is seen under storage conditions. Strips packed with a sealing film containing desiccant retain similar performance as strips packed with silica gel, 1x sealing film, even when using a sealing film 10 times or more. Therefore, the sealing film containing the desiccant can maintain the test sensitivity and shows excellent storage property under storage conditions of 1 month or more.

Example 3 HIV Chromatography Immunoassay Device Long Term Stability

Except as appropriately modified to detect HIV antibodies, for example, the labeling means used are commercially available polyclonal or monoclonal antibodies directed against heavy and / or light chain human IgG and the detecting means used are HIV antigens. A chromatographic strip is prepared in a similar manner to Example 2.A. above. 22 strips were packaged in aluminum bags with 5 g (A) of silica gel as desiccant, 2.2 g (B) of silica gel was packed in aluminum bag as desiccant, packaged in aluminum bags without desiccant (C), PET, 7 μm of 12 μm It is applied to a substrate consisting of a peelable layer made of Al of 1 μm, 188 μm of PET, 12 μm of white PET and blended PE, and containing a desiccant 25 g / m ² (D) or 50 g / m ² (E). Cover with sealing film of Sample 1 or 3 (Table 1). Strips, silica gel and sealing films used are exposed to 65% relative humidity at 27 ° C. for 6 hours before use. The strip packed under the five conditions mentioned is then heat sealed.

After the strips are heat sealed under the packaging conditions described above, they are stored in a 25 ° C. incubator for one day and then transferred to a 30 ° C. incubator, a 45 ° C. incubator or a 45 ° C. controlled box with 65% relative humidity. Strips are removed and tested after 0.5, 1, 3 and 6 months of storage.

As in Example 2.B., human serum from HIV-1 and HIV-2 infected individuals using normal human serum / plasma as a negative control and containing HIV-1 or HIV-2 antibodies, respectively, as positive samples Do this twice with plasma. A series of 2-fold dilutions are prepared from HIV-1 and HIV-2 samples and tested with 5 2-fold dilutions ranging from 2 ¹¹ to 2 ¹⁵ for HIV-1 and 2 ¹⁰ to 2 ^{14 for} HIV-2. . The results were read 15 minutes after the sample was added and are shown in Tables 6, 7 and 8 as the most diluted HIV sample dilution with which the signal can be visually observed. All negative control samples provide negative results for all strips under all packaging and storage conditions.

Time in months Test sensitivity after storage at 30 ° C (2 ^× diluent) HIV-1 HIV-2 0 0.5 One 3 6 0 0.5 One 3 6 Pkg 13 13 13 13 13/14 12 12 12 12 12 A B 13 13 13/14 ND ND 12 12 12 ND ND C 13 13 12 ND ND 12 12 11 ND ND D 13 13/14 13 13 13/14 12 12 12 12 12 E 13 13 13 13 14 12 12 12 12 12

(ND = not measured)

Time in months Test sensitivity after storage at 45 ° C (2 ^× diluent) HIV-1 HIV-2 0 0.5 One 3 6 0 0.5 One 3 6 Pkg 13 13 13 13 13/14 12 12 12 12 12 A B 13 13 13 ND ND 12 12 11 ND ND C 13 <11 <11 ND ND 12 <10 <10 ND ND D 13 13 13 13 13/14 12 12 12 12 12 E 13 13 13 13 14 12 12 12 12 12

(ND = not measured)

Time in months Test sensitivity after storage at 45 ° C / 65% relative humidity (2 ^× diluent) HIV-1 HIV-2 0 0.5 One 3 6 0 0.5 One 3 6 Pkg 13 13 13 13 13 12 12 12 12 12 A B 13 13 13 ND ND 12 12 12 ND ND C ND ND ND ND ND ND ND ND ND ND D 13 13/14 13 13 13 12 12 12 12 12 E 13 13 13 13 14 12 12 12 12 12

(ND = not measured)

Under all the conditions tested, the chromatographic strips packed without any desiccant (C) show the poorest performance with loss of sensitivity over time of storage. Strips packed with a higher amount of silica gel desiccant (5 g) or a sealant film containing desiccant retained performance for at least 6 months under all conditions tested. Both amounts of desiccant used in the sealing film are equally effective. Therefore, the sealing film containing the desiccant can maintain the test sensitivity of the chromatographic analyzer stored therein, and exhibits excellent performance and stability under long-term storage conditions.

While the invention has been described in various aspects, each modifications may be made to those skilled in the art without departing from the true spirit and scope of the invention as described in the specification and the appended claims.

Claims (11)

(1) at least one chromatographic strip having at least a sample addition site and a detection site, located on a substrate comprising a flat plate with or without a strip support spaced from each other and between them; (2) a sealing film is placed on the chromatography strip with or without a protective laminate placed between them to cover the chromatography strip as a whole; (3) a substrate and a sealing film portion are attached around each chromatography strip to seal each chromatography strip, (4) adhesion between the sealing film and the substrate allows the sealing film to be easily peeled from the substrate at least at the sample addition site, (5) (i) the sealing film or substrate comprises one or both of a dehumidifying agent containing film and an oxygen absorbent containing film, and (ii) the sealing film comprises a desiccant containing film and an oxygen absorbent containing film, while the other substrate is a film. And (iii) the sealing film and the substrate comprise a desiccant containing film and an oxygen absorbent containing film, while the other includes both a desiccant containing film and an oxygen absorbent containing film, or (iv) both the sealing film and the substrate. One or both of a desiccant containing film and an oxygen absorbent containing film, (6) in the case where the sealing film and / or the substrate contains a desiccant-containing film, the sealing film and / or the substrate are substantially waterproof at a portion which is not at least attached to the sealing film and / or the substrate, and the sealing film and / or the substrate contains an oxygen absorbent-containing film. In the case, the immunochromatographic analysis apparatus which is substantially oxygen impermeable at the portion other than the portion where the sealing film and the substrate are attached. The immunochromatographic assay device of claim 1, wherein the chromatography strip further has a labeling site. The immunochromatographic analysis device according to claim 1 or 2, wherein the sealing film is a multilayer film including a metal film. The immunochromatographic analyzer according to any one of claims 1 to 3, wherein the substrate is a multilayer film comprising a metal film. The immunochromatographic analysis device according to claim 3 or 4, wherein the metal film is aluminum foil. The immunochromatographic assay device according to any one of claims 1 to 5, wherein 5 to 12 chromatography strips are located on a substrate. The immunochromatographic analyzer according to any one of claims 1 to 6, wherein the sealing film and the substrate are attached by heat sealing. Two generally flat materials sealed together such that the two flat materials are easily or partially separated so that at least one portion of the capillary flow sheet is exposed, wherein the at least one flat material is a desiccant containing film or The action of water or oxygen, including a sheet material capable of supporting a capillary flow that immobilizes a biological reagent capable of reacting with a specific reaction partner under ambient conditions, wrapped between an oxygen absorbent containing film or both) An immunochromatographic assay device resistant to environmental degradation by The assay device of claim 8, wherein the at least one flat material transports at least the desiccant containing film and both flat materials are substantially waterproof at least in the portion where they are not attached to each other. The assay device of claim 9, wherein a labeled biological reagent capable of reacting with a specific reaction partner under ambient conditions is placed in a capillary flow sheet to be transported along a liquid sample that is moved along the flow sheet by capillary action. The assay device of claim 10, wherein the label is a colored material that becomes visible when a sufficient amount accumulates in a fixed biological reagent as a result of the assay run.