JPH01224662A - Method and kit for measuring methane phetamine - Google Patents

Abstract

PURPOSE:To easily measure methane phetamine (MA) with high accuracy by bringing the MA-conjugated high-polymer protein immobilized to a solid phase base and the MA in a measuring sample into competitive reaction with a reagent formed by labeling the monoclonal antibody having the extremely high specific immune reactivity with the MA. CONSTITUTION:The MA immobilized via the high-polymer protein to the solid phase base and the reagent formed by labeling the monoclonal antibody having the extremely high specific immune reactivity with the MA with the enzyme are brought into reaction with each other. Bovine serum albumin, egg white albumin, etc., are used as the high-polymer protein in this embodiment. The conjugation of the MA and the high-polymer protein is executed by a method of conjugating the MA and the high-polymer protein by 1-ethyl-3-(3- dimethylaminopropyl)carbodiimide hydrochloride, etc. Enzymes of a redox system, etc., such as peroxidase are used as the enzyme for labeling the antibody. Further, a plate for immunoassay, etc., are used as the shape of the solid phase base and polyethylene, etc., are used as the material thereof.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Field of Application] The present invention relates to a method and kit for measuring methamphetamine (hereinafter abbreviated as MA).

[Conventional technology]

With the increase in stimulant drug offenses, there is an increasing need to easily, quickly, and accurately identify people who abuse stimulants such as MA.

Currently, many methods have been proposed for detecting MA.

A method using gas chromatography or gas chromatography/mass spectrometry is considered to be an excellent method for accurately quantifying minute amounts of MA. However, these methods require the sample used for measurement to be pretreated by solvent extraction or adsorption chromatography, making the operations up to measurement complicated and time-consuming. There is.

Although the method using the Simon reaction is a reaction specific to secondary amines, there is a problem in that the specificity for MA is low in order to measure only MA.

Many methods using antiserum against MA have been reported so far [for example, K, Aoki et al.; J, Ph.
arm, Dyn, 6.33 (1983), S. To
kura; Anal, Bi.

chem, 161.117 (1987)), but none of these have sufficient specificity for MA, especially
There is a problem in that it has high cross-reactivity with ephedrine and methylephedrine contained in cold medicines.

An excellent method to solve these problems is considered to be an immunoassay method using monoclonal antibodies that show very high specific reactivity to MA; To date, there have been no reports regarding a method or kit for measuring MA using a monoclonal antibody that has very high specific immunoreactivity against methamphetamine.

[Problems to be solved by the invention]

An object of the present invention is to provide a method and method for measuring MA using a monoclonal antibody that has very high specific immunoreactivity to MA, which is a stimulant.

[Means for solving problems]

As a result of intensive research to solve the above problems, the present inventors discovered that MA can be easily and highly sensitively measured by measuring MA using an MA measurement kit, and have developed the present invention. It has been completed.

That is, the present invention provides the following methods: (1) A methamphetamine-binding polymer protein immobilized on a solid support and methamphetamine in a measurement sample are combined with an enzyme using a monoclonal antibody that has very high specific immunoreactivity for methamphetamine. and (2) a method for measuring methamphetamine, which is characterized by competitively reacting with a reagent labeled with (a) methamphetamine-binding polymeric protein and (b) methamphetamine. The present invention relates to a methamphetamine measurement kit characterized in that it requires a reagent in which a monoclonal antibody having extremely high specific immunoreactivity is labeled with an enzyme.

The present invention will be explained in detail below.

The MA measurement kit used in the MA measurement method of the present invention uses an enzyme to prepare a monoclonal antibody that has extremely high specific immunoreactivity for methamphetamine-binding polymer protein (hereinafter abbreviated as rMA-binding protein 1) and methamphetamine. For the measurement of 0MA, which requires a labeled reagent (hereinafter abbreviated as r-enzyme labeled antibody 1), in addition to these reagents, a solid phase support, a washing solution, a blocking solution,
Since an MA solution and an enzyme substrate solution are also required for creating an MA calibration curve, these may be incorporated into the MA measurement kit in advance, or may be prepared before the measurement.

The preparation of fMA binding protein iL of the MA measurement kit of the present invention has very high specific immunoreactivity towards MA immobilized on a solid support via a polymer protein and MA. This is necessary for reacting a monoclonal antibody with an enzyme-labeled reagent. The polymeric proteins used for its production include bovine serum albumin (BSA), ovalbumin (OVA), and Jinkasagu hemocyanin (KLH).
), T-globulin, etc.

The method for binding MA and a polymeric protein is to combine MA and any of the above-mentioned polymeric proteins with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (hereinafter referred to as EDP).
(abbreviated as C), 1-cyclohexyl-3-(2-molar ethyl)carbodiimidemeth-P-)luenesulfonate (hereinafter abbreviated as CMEC), and the like. IFMA binding protein j
However, in order to further increase the sensitivity of MA measurement, 5ephadexSSe
It is preferable to use it after purifying it by gel filtration using phacryl or the like.

The rMA-binding protein j fraction obtained by such gel filtration should be used as is (if the protein concentration is low, it should be concentrated to a specified concentration using a dialysis membrane, etc.; if the protein concentration is high, it should be diluted to a specified concentration). However, it can be dialyzed with a buffer solution adjusted to a pH around neutrality (e.g., phosphate buffer or Tris-HCl buffer), and stored by lyophilization or filtration with a sterilizing filter, and as needed. 'M
It is preferable to use the A-binding protein solution at a protein concentration of 0.1 to 100 μg/ml, preferably 1 to 20 μg/ml.

Furthermore, if necessary, a required amount of a protein preservative such as sodium azide can be added to the rMA binding protein.

(vi) In the preparation of enzyme-labeled antibody 1, enzymes for labeling the antibody include peroxidase, catalase, glucose oxidase, lactate oxidase, alcohol oxidase, monoamine A small number of enzymes selected from redox enzymes such as oxidase and β-galactosidase, and phosphate hydrolases such as alkaline phosphatase.
Both can include one or more enzymes.

Antibodies labeled with the above-mentioned enzymes are disclosed in Japanese Patent Application No. 62-2
MA-, a hybridoma strain shown in No. 53781.
7 strains (Feikoken Joyori No. 1494), MA-13 strain (Feikoken Joyori No. 1495), MA-15 strain (Feikoken Joyori No. 1)
496) can be mentioned.

The "enzyme-labeled antibody" of the present invention can be obtained by a one-step method [Immunochemistry (Imm.
Unochemistry), 6.43 (1969
)) or two-step method [Immunochemistry (Immun
chemistry), 8.1175 (197
1)), periodic acid oxidation method [Methods in Engymoro
gy), 1 Ko, 133 (1975)) and the maleimide method [Journal of Biochemistry (Jou
RNA of the Biochemist
78, 235 (1975)), but it is preferable to use the latter two methods.

This can be used as is to measure MA, but M
To further increase the measurement sensitivity of A, use 5ephadex.
It is preferable to use r-enzyme labeled antibody 1 obtained by purifying it by gel filtration using SSephacryl or the like as a reagent for measuring MA.

One portion of the r-enzyme-labeled antibody obtained by such gel filtration can be used as is, but it can be used in a buffer solution (e.g., phosphate buffer or Tris-based
Hydrochloric acid buffer, etc.), freeze-dry or filter with a sterilization filter, and store, if necessary, as a single solution of r-enzyme-labeled antibody at 0.01 to 100 μg/m/! The protein concentration is preferably 0.1 to 10 μg/m 12 .

The solid phase support used in the MA measurement method of the present invention is necessary as a support for immobilizing the rMA binding protein t1 of the MA measurement kit.

Examples of the shape of the solid phase support that can be used to immobilize the MA-bound polymer occlusal white matter include immunoassay plates, tubes, beads, membranes, etc., and the materials thereof include, for example. , polyethylene, polystyrene, polypropylene, nitrocellulose,
Examples include glass.

The washing solution used in the MA measurement method of the present invention immobilizes a certain amount of rMA-bound opercular white matter 1 on a solid support and then washes and removes what is not immobilized on the solid support.
Necessary for washing and removing unreacted substances after competitively reacting MA in the measurement sample with rMA-binding protein 1 immobilized on a solid support with r-enzyme-labeled antibody 1. .

The cleaning liquid is also used as a diluent for a measurement sample or as a solvent when preparing a blower kinder solution.

Examples of cleaning liquids used for this purpose include water,
Add Twee to the buffer solution (phosphate buffer, Tris-HCl buffer, etc.) adjusted to the pH during the reaction.
n20. (containing a surfactant such as Tween 60 in an amount of 0 to 3% by volume), but it is preferable to use a surfactant containing 0.02 to 0.8% by volume of the above-mentioned surfactant during the reaction.
It is preferable to use a buffer solution adjusted to H.

What is the blocking solution used in the MA measurement method of the present invention? This is necessary to prevent the enzyme-labeled antibody 1 from non-specifically binding to the surface of the solid support on which the rMA-binding protein 1 is not immobilized.

Blocking solutions used for this purpose include BSA, O
It can be prepared by dissolving polymeric pallidum such as VA and KLH1γ-globulin in a washing solution adjusted to the pH during the reaction, but the concentration of these polymeric proteins is 0.1 to 10 wt/vol. %, preferably O
, L ~ 2 wt/vol 1%, and when preparing using the serum of various animals, use the above-mentioned washing solution to adjust the concentration to 1-50 vol/vol 1%, preferably 10-50 vol/vol 1%.
It is preferable to set the amount to 20 vol/vol1%.

Further, if necessary, a required amount of a protein preservative such as sodium azide, sodium ethylmercury thiosalcylate, etc. can be added to the blocking solution.

As the measurement sample used in the MA measurement method of the present invention, human body fluids such as human urine, blood, and serum can be used.

In the measurement of MA, these measurement samples are diluted with the above-mentioned washing liquid within a range where MA can be measured.

Enzyme substrate solution (hereinafter referred to as r) used in the MA measurement method of the present invention
(abbreviated as "substrate solution") is "a substrate for H2C, with which the enzyme used for labeling the antibody in enzyme-labeled antibody j reacts;
and 0-phenylenediamine, 2,2"-aminobis(3
Ethylbenzothiazoline-6-sulfonic acid (ABTS)
A buffer solution containing substances such as can be used.

As described above, the solid phase support, washing solution, blocking solution, MA solution for creating a MA calibration curve, and V substrate solution j were prepared, and the MA measurement kit fMA binding protein 1,
Using an "enzyme-labeled antibody", MA in a measurement sample can be measured through the following steps.

(2) Step of immobilizing a fixed amount of rMA-binding protein on the solid support A fixed volume of the rMA-binding protein solution is brought into contact with the solid support for a fixed period of time. The temperature at the time of contact is the rMA binding protein.
Although there is no particular limitation as long as the solution does not freeze or boil, the temperature is preferably 2 to 40°C.

■ rMA-binding protein 1 not immobilized on a solid support
After contacting the solid phase support with the rMA binding protein solution for a certain period of time, remove the rMA binding protein 1 solution, and further remove the remaining rMA binding protein j without being immobilized on the solid phase support with a washing solution. Remove by washing several times.

■ Step of preventing non-specific binding of the r-enzyme-labeled antibody 1 to the surface of the solid phase support on which the rMA-binding protein 1 is not immobilized. Contact with blocking solution for a certain period of time. The temperature at the time of contact is not particularly limited as long as the blocking liquid does not freeze or boil, but is preferably 2 to 40°C.

■MA in the measurement sample and rMA immobilized on the solid support
Step 1: Competitively react binding protein j with enzyme-labeled antibody 1. Take equal volumes of the measurement sample and r enzyme-labeled antibody 1 solution,
A contact reaction is carried out with the rMA-binding protein j immobilized on the solid support. The measurement sample and the r-enzyme labeled antibody 1 solution are:
Immediately after mixing, contact reaction may be carried out with "rMA binding protein immobilized on a solid phase support," or, alternatively, each may be sequentially contacted with "rMA binding protein immobilized on a solid phase support, and then immediately stirred." It is also possible to react by

■ Step of removing the r-enzyme-labeled antibody that is not immobilized on the solid support via the rMA-binding protein J. The mixture of the measurement sample and the r-enzyme-labeled antibody solution is removed, and the rMA
The remaining measurement sample that is not bound to the solid support via the binding protein 1 is removed by washing the mixture of the enzyme-labeled antibody 1 solution several times with a washing solution.

(2) A step of reacting the r-enzyme-labeled antibody j immobilized on the solid support via the rMA-binding protein 1 with the r-substrate solution.The r-substrate solution J used here is the enzyme-labeled antibody
Add a substance that corresponds to the enzyme used to label the antibody (including a substance that changes color when an enzymatic reaction occurs).

A certain amount of the same is reacted with the r-enzyme-labeled antibody J immobilized on a solid support via the rMA-binding protein J for a certain period of time, preferably using an acid such as HzSOi, an alkali such as NaOH, or an enzyme inhibitor. It is better to stop the enzymatic reaction. There is no particular problem with the reaction temperature as long as it is within the optimum temperature range of the enzyme used at that time, but it is preferably 20 to 3
5°C is good.

(2) Measuring the absorbance of the reaction solution after the enzymatic reaction The absorbance of the reaction solution after the enzymatic reaction is measured at the wavelength at which the coloration of the reaction solution shows the maximum absorbance.

As described above, a calibration curve is created from the results of using a known amount of MA in place of the measurement sample, and the MAi in the measurement sample is
I can know i.

``r-enzyme-labeled antibody'' in this MA measurement procedure
At the step of adding the solution and the measurement sample, the MA of the i'MA-binding protein immobilized on the solid support and the MA in the measurement sample are
By the competitive reaction of r-enzyme-labeled antibody j against , the amount of MA in the measurement sample can be measured quickly and with high sensitivity.

Below, the creation of rMA-conjugated blank t,+, and the production and purification of antibodies are shown as reference examples.

Reference example 1 rMAP person white-benzene 12m
2, dissolve 0.5 mg of MA, and add 1.16 mg to this.
m g of N-(4-bromobutyl)phthalimide and 0.54 g of sodium carbonate were added and heated at 80°C for 32 hours.
After refluxing for a period of time to remove inorganic substances, 12 mf of IN hydrochloric acid was added, and the aqueous layer was washed with benzene. This aqueous layer was adjusted to pH 10 using sodium hydroxide, chloroform was added, and the resulting chloroform layer was washed with saturated brine, dehydrated over anhydrous sodium sulfate, and chloroform was distilled off under reduced pressure. This residue was dissolved in 12 m2 of ethanol, and 60 μ2 of 90% hydrazine hydrate was added thereto for 7 days at 5°C.
After refluxing for 2 hours, ethanol was distilled off. This residue was dissolved in 12 ml of IN hydrochloric acid and washed with chloroform. This was adjusted to pH 10 using sodium hydroxide, chloroform was added, and the resulting chloroform layer was washed with saturated brine, dehydrated over anhydrous sodium sulfate, and chloroform was distilled off under reduced pressure. In this way, 0.2 g of N-(4-aminobutyl)methamphetamine (hereinafter referred to as A
(abbreviated as BMA) was obtained.

"rMA binding protein" was prepared using ABMA as described below.

An aqueous solution of 20 mg ABMA and bovine serum albumin (BSA) in 0.4 m 12 dimethylformamide (20
mg/mf), add 1 mf of 10% EDPC, adjust the pH to 5.5, stir for 13 hours at room temperature and shield from light, then dialyze against pure water, and freeze-dry the non-dialyzed fraction. Thus, 22 mg of a combination of MA and BSA (hereinafter abbreviated as MA-BSA) was obtained.

Reference example 2: Toyosato Kamiyosato with a small body Made by the method described in Japanese Patent Application No. 1983-253781.

The monoclonal antibody produced by the present inventors and highly specific to MA was produced as follows.

B A of 107 cell lines suspended in phosphate buffer
The test was performed by intraperitoneally administering pristane to LB/c mice (male, 8 weeks old, 0.5 ml of pristane was administered intracavitally 2 weeks earlier). A remarkable increase in mouse body weight was observed from around the first week, and ascites was removed as appropriate from the first to third weeks. The antibody titer of the monoclonal antibody thus obtained was 106 to 1.
It was 0".

Purification of monoclonal antibodies from the obtained ascites was performed as follows.

The ascites was dialyzed against Tris-HCl buffer (pH 7.4) and applied to a DEAE-cellulose column equilibrated with the same buffer. The pass-through fraction was salted out with 50% saturated ammonium sulfate, and the resulting precipitate was dissolved in phosphate buffer (pH 7.4) and dialyzed against the same buffer.

The purity of the monoclonal antibodies against MA thus obtained was determined by slab gel electrophoresis using SDS polyacrylamide gel, and it was found that all antibodies were highly pure.

By the MA measurement method (ELISA method) using the purified monoclonal antibody against MA obtained as above,
It has become possible to measure MA with high sensitivity and quickly.

〔Example] Examples of the present invention will be specifically described below.

Note that these Examples are for illustrating the present invention, and do not limit the scope of the present invention.

The MA measurement kit fMA-binding Fluorescent Sho 1 and the "enzyme-labeled antibody" were produced as follows.

rMA-binding protein, which was obtained as shown in Reference Example 1, was dialyzed against 0.1 M phosphate buffer (pH 7,4) at 4°C overnight, and 0.2 The solution was dispensed in ml portions and lyophilized to preserve the content (5 to 100 μg/bottle), which was used as the lrMA binding protein measurement kit of the present invention. M
When measuring A, add 0.2 ml of distilled water to this bottle.
In addition, the freeze-dried powder is well dissolved and 0.0% is added to the desired concentration.
It was used after diluting with 1 M phosphate buffer (pH 7,4).

The antibody in "r enzyme-labeled antibody" was obtained as a purified monoclonal antibody as shown in Reference Example 2 using MA-15 strain (Feikoken Jokyo No. 1496).

Using this monoclonal antibody, the antibody was then labeled with an enzyme by a known method such as Shimezu to produce r-enzyme-labeled antibody 1 of the assay kit of the present invention.

First, 7.32 mg of horseradish peroxidase was dissolved in 1 m2 of distilled water, and 0.1 M sodium periodate was dissolved in 2 m2 of distilled water.
00 μ2 was added thereto and left at room temperature for 30 minutes. This enzyme solution was dialyzed overnight at 4°C using 1mM acetate buffer (pH 4.5), then 0.2M sodium carbonate buffer (pH 9.5).
100μ2 was added to adjust the pH to 9.5. on the other hand,
8 dissolved in 0.1M phosphate buffer (pH 7.4)
0.mg of monoclonal antibody (MA-15). OI
4° using M sodium carbonate buffer (PH9,5)
Dialyzed overnight at C. The peroxidase and monoclonal antibody thus obtained were mixed and allowed to stand at room temperature for 2 and a half hours, and sodium tetrahydroborate was added to this reaction solution and allowed to stand at 4"C for 2 hours. The obtained peroxidase-labeled monoclonal antibody was dialyzed overnight at 4°C using 0.1M phosphate buffer (pH 7.4), and then dialyzed against 0.1M phosphate buffer (pH 7.4) containing 0.1% sodium azide.
Dilute 100 times with M phosphate buffer (pH 7,4), dispense 0.2 ml into bottles, freeze-dry and store, and prepare the MA measurement kit ``r enzyme-labeled antibody'' (10 μl).
g/bottle). When measuring MA, add 0.2 ml of distilled water to this bottle to thoroughly dissolve the freeze-dried powder.
It was used after being appropriately diluted with 0.1 M U acid buffer (pH 7.4).

'MllW white matter a MA-BSA (MA and BS
MA-BSA (conjugate with A, 5 μg/ml) was dispensed in LOOul portions into a 96-well flat bottom plate for immunoassay (Nunc polystyrene microplate), which is a solid phase support, and incubated overnight at 4°C. was immobilized in each well. Next, to prevent nonspecific adsorption of enzyme-labeled antibody 1, a washing solution containing 10% bovine serum (pH 7.4 containing 0.05% Tween 20) was added to the plate. The urine of a healthy person (HYLAND DIAGNO
3TIC) was made 5 times more with the same cleaning solution.
2 standard MA solution [(0-15 ng) 150 μf) and 50 uf of the enzyme-labeled antibody J solution prepared in Example 1 were added to each well at the same time, stirred, and allowed to stand at room temperature for 30 minutes. After washing with washing solution, rsubstrate solution J (20 mg
of O-phenylenediamine and 10μ2 of 35% HzO2
(dissolved in 25 m2 of 0.1 M citrate buffer at pH 5.0) was dispensed into 100 μ2 portions and allowed to stand at room temperature for 5 minutes in the dark. Finally, the enzyme reaction was stopped by further dispensing 50 μλ of 2N sulfuric acid, and the absorbance at 492 nm of the solution after the reaction was stopped was measured using a microplate photometer. As a result, "this M using enzyme-labeled antibody j"
For the measurement method of A, MA could be measured with high sensitivity in about 40 minutes using a 96-well flat bottom plate for immunoassay (polystyrene microplate manufactured by Nunc) on which [mMA binding protein] was immobilized. The results are shown in FIG.

Example 3 11. of MA. M in the urine of a healthy person instead of the urine of a healthy person in Example 2
MA was measured in the same manner as in Example 2, except that a solution containing A (a solution containing MA at 30 ng/mA) was used. As a result, the added MAil and the actually measured amount of MA almost matched. The results are shown in Table 1.

Example 4 Adding MA B - Instead of A healthy person's urine j in Example 2, r Add M to healthy person's urine.
MA was measured in the same manner as in Example 2, except that solution 1 to which A was added (100 ng/mj!, solution to which MA was added) was used. As a result, the amount of MA added and the amount of MA actually measured almost matched. The results are shown in Table 1.

Example 5 71. of MA. M in the urine of a healthy person instead of the FR regular-sized urine of Example 2
A solution containing A (M to be 300 ng/ml!)
MA was measured in the same manner as in Example 2, except that the solution containing A) J was used. As a result, the added MAfi and the actually measured amount of MA almost matched. The results are shown in Table 1.

Example 6 Addition of MA B - Add M to r healthy person's urine instead of r healthy person's urine 1 in Example 2
A solution containing A (M to be 1000 ng/mf)
MA was measured in the same manner as in Example 2, except that the solution containing A) 1 was used. As a result, the amount of MA added and the amount of MA actually measured almost matched. The result is the first
Shown in the table.

Example 7 MA'7 Instead of 1 of Example 2, 1 of healthy person's urine 1, ``M
MA was measured in the same manner as in Example 2, except that solution 1 to which A was added (a solution to which MA was added to give 3000 ng/mj!) was used. As a result, the amount of MA added and the amount of MA actually measured almost matched. The results are shown in Table 1.

(Hereinafter, blank space) In Table 1 Example 2, instead of r healthy person's urine 1, r healthy person's urine was used.
A solution to which A and methylephedrine (hereinafter abbreviated as ME) were added (MA warm solution of 0.5 μg/rr#t, solution to which M amounts of 0.1' and 10.1100 t1/ml were added, respectively) 1 was used. Except for this, MA was measured in the same manner as in Example 2. As a result, the amount of MA added (0.5μg/mf
) and the amount of MA actually measured (0.35~0.55 u
g/ml) were almost the same. The results are shown in Table 2.

Example 9 Overflow of MA in ME R - Parallel Example 2: M in the urine of a healthy person instead of "urine of a healthy person"
A solution containing A and ME (1,0 μg/ml MA warm solution, 100 μg/ml of ME at 0, 1, 10°, respectively)
MA was measured in the same manner as in Example 2, except that the solution containing m 1 added) 1 was used. As a result, the amount of MA added (1,0 μg/mf) and the amount of MA actually measured (1,2
2 to 1.38 tt g/mf) were almost in agreement.

The results are shown in Table 2.

Example 10 Adding MA to ME Kuchiwipe Ogi Example 2: Adding M to the urine of a healthy person instead of the urine of a healthy person
A and ME added solution (5,0 μg/m℃ MA
MA was measured in the same manner as in Example 2, except that a warm solution and a solution to which ME was added at 0, 1, 10:1100 u/mf, respectively) were used. As a result, the amount of MA added (5, O
ug/ml”) and the amount of MA actually measured (5,37~5
．． 78μg/m! ! , ) were almost in agreement. The results are shown in Table 2.

Table 2 According to Example 2, in addition to MA, amphetamine (AP
), methylephedrine (ME), and methoxyphenamine (MPA) were measured. The results are shown in Table 3 and FIG.

Table 3 [Effects of the Invention] According to the present invention, by using an MA measurement kit that requires rMA-binding protein and enzyme-labeled antibody J, MA can be measured rapidly and with high sensitivity under competitive reaction conditions. It can be measured by

[Brief explanation of the drawing]

FIG. 1 shows an MA standard curve prepared using r-enzyme-labeled antibody 1. Figure 2 shows MA, amphetamine (AP), methylephedrine (ME),
The results of examining the reactivity of methoxyphenamine (M, PA) are shown. Patent applicant Ube Industries, Ltd. Figure 1 Methamphetamine concentration (ng/mj)

Claims (2)

[Claims] (1) A methamphetamine-binding macromolecular protein immobilized on a solid support and methamphetamine in the measurement sample compete with a reagent labeled with an enzyme and a monoclonal antibody that has very high specific immunoreactivity for methamphetamine. A method for measuring methamphetamine that is characterized by a reaction. (2) A reagent used in a method for measuring methamphetamine, which is an enzyme-labeled monoclonal antibody that has (a) a methamphetamine-binding polymeric protein and (b) a monoclonal antibody that has extremely high specific immunoreactivity for methamphetamine. A methamphetamine measurement kit characterized by: