JP2011021145A - Treating material for explosive and method for removing the explosive - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method for removing an explosive, in which an antibody having bondability to acetone peroxide to be removed, particularly trimeric acetone peroxide, is newly created, the trimeric acetone peroxide is bonded to the immobilized antibody by applying an antigen-antibody reaction by utilizing the created antibody, and the trimeric acetone peroxide bonded to the antibody is recovered, wiped out completely and removed; and to provide a wiping material which is utilized in the wiping/removing work and to which the antibody is immobilized. <P>SOLUTION: A monoclonal antibody exhibiting cross reactivity to the acetone peroxide is selected from monoclonal antibodies to 3-[12-(2-carboxyethyl)-9,12-dimethyl-7,8,10,11,13,14-hexaoxa-spiro-[5.8]tetradec-9-yl]-propanoic acid having the structure similar to that of the acetone peroxide. The acetone peroxide is bonded to the selected and immobilized monoclonal antibody and the acetone peroxide bonded to the antibody is recovered, wiped out completely and removed. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to a method for recovering and treating a peroxide derivative by using a monoclonal antibody capable of binding to a peroxide derivative, which is used as an explosive, and for wiping and removing it safely, as well as the wiping and removing work. The present invention relates to a wiping treatment material used in the field.

Organic compounds used as explosives, for example, TNT (2,4,6-trinitrotoluene), RDX (hexogen; 1,3,5-trinitro-1,3,5-triazinan), pen slit (pentaeritetranitrate) Slit: C (CH ₂ ONO ₂ ) ₄ ) has a nitro group in its molecule. Nitro compounds, which are widely used as explosives, have relatively low friction sensitivity and impact sensitivity.

On the other hand, organic peroxide derivatives used as explosives have relatively high friction sensitivity and impact sensitivity. Therefore, organic peroxide derivative explosives are dangerous substances that easily explode when subjected to friction or impact. In fact, organic peroxide derivative explosives, acetone peroxide, especially trimeric TATP (C ₉ H ₁₈ O ₆ : triacetone triperoxide) or hexamethylenetriperoxidediamine (HMTD) It can be used as a primary explosive because it is very sensitive to friction and shock.

Among them, acetone peroxide, in particular, trimer type TATP (C ₉ H ₁₈ O ₆ : triacetone triperoxide) having a structure represented by the following formula (I) is a relatively easily available raw material. Synthesis can be performed on a laboratory scale using acetone and hydrogen peroxide, and sulfuric acid and hydrochloric acid, which are used as acid catalysts. In addition, it has good crystallinity and can be isolated and purified as white crystals having a melting point of 91 ° C.

  Trimer-type TATP is sometimes used in explosive terrorism because of the ease of production.

  Trimeric TATP crystals that have been isolated and purified are extremely sensitive to friction and impact, so there is a risk of explosion when collecting and processing trace amounts of crystals. is there. On the other hand, trimer-type TATP, when dissolved or soaked in an organic solvent such as acetonitrile, drastically lowers the friction sensitivity and impact sensitivity, so that the organic solvent is soaked when the crystal is recovered and processed. A technique of wiping with an absorbent material such as a sponge is used. In the method of wiping in a state of being dissolved or immersed in the organic solvent, a small amount of the liquid in which the trimer-type TATP is dissolved remains, but remains. When the organic solvent contained in the remaining liquid evaporates, a fine powder of trimer-type TATP is deposited. For this reason, it is difficult to completely recover and remove the trimer-type TATP by a method of wiping after dissolving or immersing in an organic solvent.

JP 2009-23985 A

Chemistry Letters, Vol.35, No.10, p.1126-1127 (2006) Organic & Biomolecular Chemistry Vol.4, p.4431-4436 (2006)

  A method of completely removing the fine powder of trimer-type TATP that inevitably remains after being collected and treated using a method of wiping after being dissolved or immersed in an organic solvent Development is required.

  For example, as a technology to completely remove or detoxify harmful substances, an antibody against influenza virus, which is a virus particle showing pathogenicity, is used to fix the virus particle by applying an antigen-antibody reaction. A method of removing and removing the image has been proposed (see Japanese Patent Application Laid-Open No. 2009-23985). The technique of immobilizing and removing the target virus particles by applying an antigen-antibody reaction is effective as a means for completely removing influenza viruses scattered in the atmosphere. If an antibody that has binding ability to the target to be removed is available, the antibody is immobilized in advance, and then an antigen-antibody reaction is performed with the target substance to be removed. Is possible.

  In order to apply the removal means applying this antigen-antibody reaction to the collection and removal of fine powdered trimeric TATP, an antibody capable of binding to the trimeric TATP is required. For example, since the organic peroxide explosive compound itself having a structure represented by the formula (I), such as a trimer type TATP, does not exhibit immunogenicity, the organic peroxide explosive compound No specific antibody has been reported so far. If an antibody having a selective binding ability to the organic peroxide-type explosive compound is available, the organic peroxide-type explosive compound is immobilized using the antibody having the selective binding ability. There is a high possibility that it can be applied to the development of a process for removing and removing a gas.

  The present invention solves the aforementioned problems. That is, an object of the present invention is to newly create an antibody having a binding ability to a target peroxide derivative-type explosive and apply the antigen-antibody reaction using the created antibody to detect the target excess. Provided is a method of completely wiping and removing an oxide derivative-type explosive by binding to an immobilized antibody and recovering the same, and a wiping treatment material on which the antibody is immobilized and used for the removal operation There is.

The object of the present invention is, for example, a peroxide derivative type explosive to be removed, acetone peroxide, in particular, a trimer type TATP (C ₉ H ₁₈ O ₆ having the structure shown in the following formula (I): : A new antibody having a binding ability to triacetone triperoxide) is applied, and the antigen-antibody reaction is applied using the created antibody to fix the trimeric TATP shown in the formula (I). An object of the present invention is to provide a method of completely wiping and removing by binding to and recovering the antibody, and a wiping treatment material immobilizing the antibody and used for the wiping and removing operation.

  TATP (3,3,6,6,9,9-hexamethyl-1,2,4,5,7,6-hexaoxacyclononane):

  In order to solve the above-mentioned problems, the present inventors first studied a technique for newly creating an antibody having a binding ability to a target peroxide derivative-type explosive.

  It has already been found that the subject peroxide derivative explosives themselves are not immunogenic. On the other hand, even a low molecular weight compound having no immunogenicity may function as an immunogen when the low molecular weight compound is bound onto a carrier protein to form a modified protein modified with the low molecular weight compound. (Non-patent document 2: Chemistry Letters, Vol. 35, No. 10, p. 1126-1127 (2006)). There are many cases in which antibodies that show specific reactivity to low molecular weight compounds that do not have immunogenicity using this technique have been produced, but they are not effective against all low molecular weight compounds. That is, in the modified protein, whether or not the site to which the low molecular weight compound is bound actually shows immunogenicity depends on the three-dimensional structure of the site, and is effective for all low molecular weight compounds. It's not that.

  However, for example, trimeric acetone peroxide (TATP) itself having the structure shown in formula (I) has a functional group that can be used to produce a modified protein by binding to a carrier protein. The above-mentioned method cannot be applied.

  Furthermore, the present inventors have noted that in an antigen-antibody reaction, an antibody may exhibit a phenomenon that exhibits reactivity with a substance having a structure similar to the original antigen, so-called cross-reactivity. did. That is, in place of the target low molecular weight compound, when many kinds of specific antibodies against an antigen having a structure similar to the low molecular weight compound are created, Thus, it was conceived that there may be an antibody showing cross-reactivity.

  In the trimeric acetone peroxide (TATP) having the structure represented by the formula (I), the present inventors actually have a characteristic structure of the ring structure. Among many types of low molecular weight compounds having similarities, the inventors searched for an immunogenicity of the resulting modified protein when bound on a carrier protein. Next, among the many types of antibodies created by immunizing mice using the modified protein as an immunogen, the cross-reactivity to the trimeric acetone peroxide (TATP) of the formula (I) A search was conducted as to whether or not there was an antibody indicating the above. When the above two-step search process was carried out, fortunately, when the compound represented by the following formula (II) was bound on the carrier protein, the resulting modified protein showed immunogenicity, It has been found that antibodies exhibiting cross-reactivity exist among many types of antibodies created by immunizing mice using the modified protein as an immunogen.

  3- [12- (2-Carboxyethyl) -9,12-dimethyl-7,8,10,11,13,14-hexaoxa-spiro- [5.8] tetradec-9-yl] -propionic acid (3- [ 12- (2-carboxyethyl) -9,12-dimethyl-7,8,10,11,13,14-hexaoxa-spiro- [5.8] tetradec-9-yl] -propanoic acid)

  Specifically, for a compound represented by formula (II) created by immunizing a mouse using a modified protein obtained by binding a dicarboxylic acid compound represented by formula (II) on a carrier protein as an immunogen. A group of hybridoma cell lines producing antibodies is prepared, and antibodies exhibiting cross-reactivity to the trimeric acetone peroxide (TATP) of formula (I) are produced from the group of hybridoma cell lines. Several hybridoma cell lines could be selected.

  The monoclonal antibodies produced by several of the selected hybridoma cell lines do not show cross-reactivity with at least the endogenous substance of the mouse used as the immunized animal, but the dicarboxylic acid compound represented by the formula (II) and the formula It was confirmed that the compound had reactivity to the trimeric acetone peroxide (TATP) of (I).

  In addition to the series of findings described above, the present inventors immobilized a monoclonal antibody produced by several of the selected hybridoma cell lines, and then trimeric acetone peroxide (TATP) of the formula (I) The trimeric acetone peroxide (TATP) of the formula (I) once bound to the monoclonal antibody when the antigen-antibody reaction was performed with the monoclonal antibody, even when the aqueous solvent used for the antigen-antibody reaction was evaporated. It was also confirmed that the trimeric acetone peroxide (TATP) (I) was stably bound to the monoclonal antibody.

  As the base material for immobilizing the monoclonal antibody, a liquid-absorbing material having a liquid-absorbing property with respect to a solvent capable of dissolving the trimeric acetone peroxide (TATP) of the formula (I) was selected and dissolved in the solvent. A solution of the trimeric acetone peroxide (TATP) of the formula (I) can be absorbed with the absorbent material. At that time, the trimeric acetone peroxide (TATP) of the formula (I) contained in the solution sucked into the liquid absorbing material comes into contact with the monoclonal antibody immobilized in the liquid absorbing material. It was also confirmed that an antigen-antibody reaction can be performed. The trimeric acetone peroxide (TATP) of the formula (I) stably bound to the monoclonal antibody in the solution is stable with the monoclonal antibody even at the stage where the solvent contained in the solution is removed by evaporation. It was also confirmed that they were bonded.

Accordingly, the solid powder of acetone peroxide of the formula (I) was dissolved in an appropriate amount of a solvent capable of dissolving the acetone peroxide of the formula (I), and made of a liquid-absorbing material having liquid absorbency with respect to the solvent Using a wiping treatment material, the liquid in which the acetone peroxide of the formula (I) is dissolved can be absorbed into the wiping treatment material and wiped off;
At that time, the wiping treatment material is configured to immobilize the monoclonal antibody having the binding ability to acetone peroxide of the formula (I) in the liquid absorption material, so that the wiping treatment material absorbs the wiping treatment material. The acetone peroxide of the formula (I) dissolved in the solution of acetone peroxide of the formula (I) that has been liquefied forms a complex with the immobilized monoclonal antibody by the antigen-antibody reaction. What you can do;
As a result, it was found that the solid powder of the target acetone peroxide of formula (I) can be wiped off.

  Furthermore, the present inventors have found that the trimeric acetone peroxide (TATP) of the above formula (I) and the characteristic structure and similarity in the trimeric acetone peroxide (TATP) of the above formula (I) In addition to the combination of monoclonal antibodies against the dicarboxylic acid compound represented by the formula (II) having the structure having the structure, the subject peroxide derivative-type explosive itself does not exhibit immunogenicity. We have also found that it is possible to create a monoclonal antibody against a low molecular weight compound having a structure similar to the characteristic structure of explosives by the same method. At that time, among the many monoclonal antibodies against low molecular weight compounds having a structure similar to the characteristic structure of the peroxide derivative type explosive, a cross-reaction with the target peroxide derivative type explosive is performed. The antibody showing the cross-reactivity to be selected is wiped off from the trimeric acetone peroxide (TATP) of the formula (I) using the antigen-antibody reaction described above. We have also found that it has the characteristics necessary for the production of processing materials.

  The present inventors have completed the present invention based on the above-described series of findings and verification results.

That is, the explosive wiping and removing method according to the present invention is:
Peroxide derivative type explosive, acetone peroxide of the following formula (I) (TATP: 3,3,6,6,9,9-hexamethyl-1,2,4,5,7,6-hexaoxacyclononane ) By wiping and removing the solid powder,

The method of wiping and removing is as follows:
Dissolving a solid powder of acetone peroxide of formula (I) in an appropriate amount of a solvent capable of dissolving the acetone peroxide of formula (I);
Using a wiping treatment material made of a liquid-absorbing material having a liquid-absorbing property with respect to the solvent, the wiping treatment material absorbs the liquid in which acetone peroxide of the formula (I) is dissolved, and is wiped off. It has a process,
The wiping treatment material is
A monoclonal antibody having binding ability to acetone peroxide of formula (I) is immobilized in the liquid-absorbing material;
The acetone peroxide of formula (I) dissolved in the solution of acetone peroxide of formula (I) absorbed in the wiping treatment material is immobilized by the antigen-antibody reaction. A method for wiping and removing explosives, characterized by forming a complex with an antibody.

in particular,
The monoclonal antibody having binding ability to acetone peroxide of formula (I) is:
A monoclonal antibody against a low molecular weight compound having a structure similar to the characteristic structure of acetone peroxide of the formula (I), and cross-reactive with the peroxide peroxide of the formula (I) It is preferable to have.

that time,
The low molecular weight compound having a structure similar to the characteristic structure of acetone peroxide of the formula (I) is a dicarboxylic acid compound having a structure represented by the following formula (II): 3- [12- (2 -Carboxyethyl) -9,12-dimethyl-7,8,10,11,13,14-hexaoxa-spiro- [5.8] tetradec-9-yl] -propionic acid (3- [12- (2-carboxyethyl) -9,12-dimethyl-7,8,10,11,13,14-hexaoxa-spiro- [5.8] tetradec-9-yl] -propanoic acid).

In the method for wiping and removing explosives according to the present invention having the above-described configuration,
The monoclonal antibody having binding ability to acetone peroxide of the formula (I) is:
A non-human mammal can be obtained by using a modified protein obtained by binding a low molecular weight compound having a structure similar to the characteristic structure of acetone peroxide of formula (I) on a carrier protein as an immunogen. A monoclonal antibody against the low molecular weight compound created by immunization,
An antibody having cross-reactivity with acetone peroxide of the formula (I) is preferable.

  In that case, it is preferable that the non-human mammal is a mouse.

  Further, in a modified protein obtained by binding the low molecular compound on a carrier protein, it is preferable to select Keyhole Limpet Hemocyanin as the carrier protein.

A compound having a carboxyl group (—COOH) in the molecule is selected as a low molecular weight compound having a structure similar to the characteristic structure of acetone peroxide of formula (I),
A modified protein obtained by binding a compound having a carboxyl group (—COOH) in the molecule onto a carrier protein comprises the carboxyl group (—COOH) and an amino group (—NH ₂ ) on the carrier protein. It is preferable that a compound having a carboxyl group (—COOH) in the molecule is bonded via an amide bond (—CO—NH—).

For example, formation of an amide bond (—CO—NH—) between the carboxyl group (—COOH) and an amino group (—NH ₂ ) on the carrier protein is performed using a carbodiimide method. Is preferred.

Furthermore, as a monoclonal antibody immobilized in the liquid-absorbing material,
A form using a monoclonal antibody produced by a hybridoma cell line: NECP-C57Z 3B-7E (FERM BP-11125) and having a structure represented by the above formula (II) is used.

In the method for wiping and removing explosives according to the present invention having the above-described configuration,
The monoclonal antibody is preferably immobilized in the liquid-absorbing material with polyvinyl alcohol.

  Or it is preferable that the said monoclonal antibody is fix | immobilized in the said liquid absorption material through the linker which consists of protein at least.

  At this time, the linker composed of at least protein is more preferably at least one of protein A and protein G.

  On the other hand, the wiping treatment material is desirably used in a step of wiping and removing after at least absorbing water or the solvent.

Furthermore, in the operation of dissolving the solid powder of acetone peroxide of the formula (I) in an appropriate amount of a solvent capable of dissolving the acetone peroxide of the formula (I),
It is preferable to dissolve the solid powder of acetone peroxide of the formula (I) by using an appropriate amount of a solvent capable of dissolving the acetone peroxide of the formula (I) to form a solution.

  In this case, the solvent capable of dissolving acetone peroxide of the formula (I) includes at least one organic solvent selected from the group of organic solvents consisting of acetonitrile, methanol, ethanol, acetone, ethanolamine, and methyl ethyl ketone. Is preferred.

  In addition, the present invention also provides an explosive wiping treatment material suitably used in the above explosive wiping removal method.

That is, the explosive wiping treatment material according to the present invention is:
Peroxide derivative type explosive, acetone peroxide of the following formula (I) (TATP: 3,3,6,6,9,9-hexamethyl-1,2,4,5,7,6-hexaoxacyclononane ) A wiping treatment material that can be used for wiping and removing solid powder,

The wiping treatment material is
Made of a liquid-absorbing material having a liquid-absorbing property to a solvent capable of dissolving acetone peroxide of formula (I)
An explosive wiping treatment material characterized in that a monoclonal antibody having binding ability to acetone peroxide of formula (I) is immobilized in the liquid-absorbing material.

in particular,
The monoclonal antibody having binding ability to acetone peroxide of formula (I) is:
A monoclonal antibody against a low molecular weight compound having a structure similar to the characteristic structure of acetone peroxide of the formula (I), and cross-reactive with the peroxide peroxide of the formula (I) It is preferable to have.

that time,
The low molecular weight compound having a structure similar to the characteristic structure of acetone peroxide of the formula (I) is a dicarboxylic acid compound having a structure represented by the following formula (II): 3- [12- (2 -Carboxyethyl) -9,12-dimethyl-7,8,10,11,13,14-hexaoxa-spiro- [5.8] tetradec-9-yl] -propionic acid (3- [12- (2-carboxyethyl) -9,12-dimethyl-7,8,10,11,13,14-hexaoxa-spiro- [5.8] tetradec-9-yl] -propanoic acid).

In the explosive wiping treatment material according to the present invention having the above-described configuration,
The monoclonal antibody having binding ability to acetone peroxide of the formula (I) is:
A non-human mammal can be obtained by using a modified protein obtained by binding a low molecular weight compound having a structure similar to the characteristic structure of acetone peroxide of formula (I) on a carrier protein as an immunogen. A monoclonal antibody against the low molecular weight compound created by immunization,
An antibody having cross-reactivity with acetone peroxide of the formula (I) is preferable.

  In that case, it is preferable that the non-human mammal is a mouse.

  Further, in a modified protein obtained by binding the low molecular compound on a carrier protein, it is preferable to select Keyhole Limpet Hemocyanin as the carrier protein.

A compound having a carboxyl group (—COOH) in the molecule is selected as a low molecular weight compound having a structure similar to the characteristic structure of acetone peroxide of formula (I),
A modified protein obtained by binding a compound having a carboxyl group (—COOH) in the molecule onto a carrier protein comprises the carboxyl group (—COOH) and an amino group (—NH ₂ ) on the carrier protein. It is preferable that a compound having a carboxyl group (—COOH) in the molecule is bonded via an amide bond (—CO—NH—).

For example, formation of an amide bond (—CO—NH—) between the carboxyl group (—COOH) and an amino group (—NH ₂ ) on the carrier protein is performed using a carbodiimide method. Is preferred.

Furthermore, as a monoclonal antibody immobilized in the liquid-absorbing material,
A form using a monoclonal antibody produced by a hybridoma cell line: NECP-C57Z 3B-7E (FERM BP-11125) and having a structure represented by the above formula (II) is used.

In the explosive wiping treatment material according to the present invention having the above-described configuration,
The monoclonal antibody is preferably immobilized in the liquid-absorbing material with polyvinyl alcohol.

  Or it is preferable that the said monoclonal antibody is fix | immobilized in the said liquid absorption material through the linker which consists of protein at least.

  At this time, the linker composed of at least protein is preferably at least one of protein A and protein G.

  The explosive wiping and removing method according to the present invention uses a monoclonal antibody against a low molecular weight compound having a structure similar to the characteristic structure of a peroxide derivative-type explosive, By selectively cross-reacting the monoclonal antibody to the peroxide derivative-type explosive, the target peroxide derivative-type explosive is selectively bound and removed through an antigen-antibody reaction. Is possible. For example, when the target peroxide derivative-type explosive is a trimeric acetone peroxide (TATP), it has a structure having a structure similar to that of the trimeric acetone peroxide (TATP). Molecular compound: 3- [12- (2-carboxyethyl) -9,12-dimethyl-7,8,10,11,13,14-hexaoxa-spiro- [5.8] tetradec-9-yl] -propionic acid ( 3- [12- (2-carboxyethyl) -9,12-dimethyl-7,8,10,11,13,14-hexaoxa-spiro- [5.8] tetradec-9-yl] -propanoic acid) The peroxide derivative-type explosive of interest via an antigen-antibody reaction by cross-reactivity of the monoclonal antibody to the peroxide-derivative-type explosive that is used and immobilized in the liquid-absorbing material Can be selectively combined and removed.

  Specifically, a solid powder of acetone peroxide of formula (I) is dissolved in an appropriate amount of a solvent capable of dissolving acetone peroxide of formula (I), and prepared with a liquid-absorbing material having liquid absorbency with respect to the solvent. Using the wiping treatment material, the liquid in which the acetone peroxide of the formula (I) is dissolved can be absorbed into the wiping treatment material and removed by wiping. In this case, the wiping treatment material is The above-described formula (I), wherein the wiping treatment material is made to absorb liquid by immobilizing the monoclonal antibody having binding ability to acetone peroxide of the formula (I) in the liquid-absorbing material. Since acetone peroxide of formula (I) dissolved in acetone peroxide solution can form a complex with the immobilized monoclonal antibody by antigen-antibody reaction, the target formula (I) It is possible to safely implement the wiping removal of the solid powder of acetone. In particular, by using the explosive wiping treatment material according to the present invention, the target peroxide derivative-type explosive: acetone solid peroxide of formula (I) can be easily wiped away. Become.

3- [12- (2-carboxyethyl) -9,12-dimethyl-7,8,10,11,13,14-hexaoxa-, which can be used in the wiping and removing method according to the first embodiment of the present invention. Spiro- [5.8] tetradec-9-yl] -propionic acid (3- [12- (2-carboxyethyl) -9,12-dimethyl-7,8,10,11,13,14-hexaoxa-spiro- [5.8 ] Graph showing the results of ELISA evaluation of the cross-reactivity of monoclonal antibodies against [tetradec-9-yl] -propanoic acid) to target peroxide derivative explosives and trimeric acetone peroxide (TATP) It is. It is a figure which shows typically the structure of the wiping off treatment material of the explosive concerning the 1st embodiment of this invention. It is a figure which shows typically the structure of the wiping off treatment material of the explosive concerning the 2nd embodiment of this invention. It is a figure which shows typically an example of the procedure which implements the wiping removal method concerning 1st embodiment of this invention. It is a figure which shows typically another example of the procedure which implements the wiping removal method concerning 1st embodiment of this invention.

  Below, the wiping removal method of the peroxide derivative type explosive concerning this invention and the wiping off processing material utilized for the wiping removal work are demonstrated in detail.

  First, an antibody having binding ability to a peroxide derivative-type explosive used in the method for wiping and removing a peroxide derivative-type explosive according to the present invention and a method for producing the antibody will be described in more detail.

  As a means of creating antibodies against low molecular weight organic compounds, if the target low molecular weight organic compound itself does not show immunogenicity, the modified carrier obtained by binding the target low molecular weight organic compound on the carrier protein There is a technique of using a protein as an immunogen (Non-patent Document 2: Chemistry Letters, Vol. 35, No. 10, p. 1126-1127 (2006)).

Specifically, a low molecular weight organic compound has a reactive functional group such as an amino group (—NH ₂ ), a hydroxyl group (—OH), a sulfanyl group (—SH), or a carboxyl group (—COOH). In such a case, the reactive functional group can be used to covalently link organic compounds having other reactive functional groups. The carrier protein has a three-dimensional structure composed of a peptide chain in which a plurality of amino acid residues are linked, but on its surface, an amino acid residue having a reactive functional group on the side chain There are several. Therefore, it has a low molecular weight that has a reactive functional group by utilizing a reactive functional group on the side chain of an amino acid residue that has a three-dimensional structure and exists on the surface of the carrier protein. It is possible to bind organic compounds. The modified carrier protein, which is modified on the surface by a low molecular weight organic compound, is a non-natural protein molecule and is recognized as a foreign substance that is different from the endogenous protein molecule of the mammal itself. Is frequent. In particular, a site that has been modified by a low molecular weight organic compound frequently exhibits immunogenicity. When the modified carrier protein surface modified with a low molecular weight organic compound exhibits immunogenicity, when the mammal is immunized with the modified carrier protein, the modified carrier Specific antibodies against proteins are created.

  There may be a plurality of sites (antigenic determinants) that exert immunogenicity on the surface of the modified carrier protein. In that case, multiple types of antibodies specific to each of the plurality of immunogenic sites (antigenic determinants) are created. Among the multiple types of antibodies that are specific to modified carriers and proteins that have been created, there are antibodies that use the low molecular weight organic compounds themselves used for their modification as sites that exhibit immunogenicity (antigenic determinants) The frequency of doing is high. By screening based on the binding ability to the low molecular weight organic compound itself used for modification, an antibody that uses the low molecular weight organic compound itself used for modification as an immunogenic site (antigenic determinant) It is possible to sort.

  However, if the mammal to be immunized already has an antibody that exhibits high cross-reactivity with the antigenic determinant present on the surface of the modified carrier protein, the antigenic determination that exhibits this cross-reactivity is determined. Creation of new antibodies against the group does not occur. That is, when an immune reaction to the modified carrier protein is possible using the high cross-reactivity exhibited by the antibody already held by the mammal to be immunized, the antigen determinant exhibiting this cross-reactivity, Creation of new antibodies does not occur.

  Furthermore, even when the modified site functions as a site that exhibits immunogenicity (antigenic determinant), the antibody specific for the antigenic determinant has a low molecular weight used for the modification. There are many cases where the binding ability to the organic compound itself is not high.

  In other words, using the technique of binding the target low molecular weight organic compound on the carrier protein and using the resulting modified carrier protein as an immunogen, it is specific to the low molecular weight organic compound itself used for modification. Whether or not a specific antibody can be created depends on the following factors. Specifically, it depends on the three-dimensional structure of the target low molecular weight organic compound itself, the combination with the carrier protein to be used, the binding form on the carrier protein, the selection of the modification site, and the above four factors. is doing.

  In fact, since the three-dimensional structure of the target low molecular weight organic compound itself has already been determined, whether or not an appropriate combination can be selected for the remaining three factors depends largely on chance. . In other words, the binding form on the carrier protein and its modification site depend on the type of carrier protein used, and are limited by the type of reactive functional group possessed by the target low molecular weight organic compound. The Depending on the three-dimensional structure of the target low molecular weight organic compound itself, an appropriate combination may not be selected for the remaining three factors.

  On the other hand, if the peroxide derivative-type explosive itself does not have a reactive functional group in its molecule, it binds the target low molecular weight organic compound onto the carrier protein described above, and obtains it. It is not possible to apply a technique that uses a modified carrier protein to be used as an immunogen. Of course, the peroxide derivative-type explosive is a low molecular weight organic compound and itself does not exhibit immunogenicity.

  Therefore, in the present invention, a large number of antibodies against low molecular weight compounds having structures having similar structures and similarities to the target peroxide derivative type explosives are created, and low molecules having structures having similarities are created. Among many antibodies against compounds, a method is employed in which antibodies having cross-reactivity with the target peroxide derivative-type explosive are selected.

  Hereinafter, as a target peroxide derivative-type explosive, trimeric acetone peroxide (TATP: 3,3,6,6,9,9-hexamethyl-1) having the structure represented by the following formula (I): , 2,4,5,7,6-hexaoxacyclononane) as an example, a method for producing an antibody used in the present invention will be described more specifically.

  The structural feature of the trimeric acetone peroxide (TATP) represented by the formula (I) is the ring structure itself. Of the low molecular weight peroxide compounds that have this ring structure characteristic and have a reactive functional group in the molecule, they are not highly toxic to mammals themselves, and when bound on a carrier protein We searched for those capable of retaining the ring structure.

  As a result of the search, a dicarboxylic acid compound having a structure represented by the following formula (II) among a group of low molecular weight peroxide type compounds that are being investigated for use as antimalarial drugs having low toxicity: 3- [12- (2-carboxyethyl) -9,12-dimethyl-7,8,10,11,13,14-hexaoxa-spiro- [5.8] tetradec-9-yl] -propionic acid (3- [12- (2-carboxyethyl) -9,12-dimethyl-7,8,10,11,13,14-hexaoxa-spiro- [5.8] tetradec-9-yl] -propanoic acid) as a candidate satisfying the above requirements ,chosen.

In the present invention, a reactive functional group present on the surface of a carrier protein is utilized by utilizing a carboxyl group (—COOH), which is a reactive functional group present in the molecule of a dicarboxylic acid compound having the structure represented by formula (II). It is preferable to select a form to be bonded to the surface of the carrier protein by forming an amide bond (—CO—NH—) with a group, particularly an amino group (—NH ₂ ).

  At that time, the carrier protein binds the target low molecular weight organic compound onto the carrier protein, and uses the resulting modified carrier protein as an immunogen (Non-patent Document 2: Chemistry Letters, Vol. 35, No. 10, p. 1126-1127 (2006)), various carrier proteins already used can be used. As the carrier protein, bovine serum albumin, bovine thyroglobulin, Keyhole Limpet Hemocyanin and the like can be suitably used.

  When a target low molecular weight organic compound is bound on a carrier protein and the resulting modified carrier protein is used as an immunogen, a specific antibody against the modified carrier protein is used in an immunized mammal. Is created. In this case, since the carrier protein itself to be used generally has immunogenicity, in addition to the antibody specific to the modified site in the modified carrier protein, it is specific to the antigenic determinant of the carrier protein itself. New antibodies are also created.

  Considering this point, it is preferable that the mixing ratio of unmodified carrier / protein is low with respect to the modified carrier / protein used for immunization. Of course, it is more preferable to use a modified carrier protein that does not contain unmodified carrier protein.

  On the other hand, on the carrier protein to be used, there are generally a plurality of sites (modifiable sites) where the target low molecular weight organic compound can be bound and modified. Each of these modifiable sites has a reactive functional group, but generally there is a difference in reactivity. Therefore, the binding of the low molecular weight organic compound of interest proceeds preferentially from the highly reactive modifiable site, and the low molecular weight organic compound of interest is consumed. , The efficiency with which the low molecular weight organic compound of interest is bound tends to be further reduced. In order to achieve binding of the low molecular weight organic compound of interest to all of the plurality of modifiable sites present on the carrier protein, the amount of the low molecular weight organic compound of interest used in the reaction must be multiple. It is desirable to select a considerably excessive amount relative to the total number of modifiable sites. For example, when there are N modifiable sites on the carrier protein, the amount of the low molecular weight organic compound to be used for the reaction requires a minimum of N molecules per carrier protein molecule. However, it is desirable to select at least 50 molecules or more, preferably 60 molecules or more.

  The immunization can be performed by, for example, administering a solution containing an effective amount of a modified carrier protein combined with a low molecular weight organic compound of the subject to a mammal to be immunized by injection. desirable. As the administration form by injection, subcutaneous injection, intradermal injection, intravenous injection, or intraperitoneal administration can be used. The solution is usually administered by subcutaneous injection. At this time, it is preferable to add various adjuvants to a solution containing an effective amount of the modified carrier protein. Usually, an adjuvant that has been conventionally used for immunization can be used as the adjuvant. Available adjuvants include Freund's complete adjuvant, water-in-oil-in-water emulsion, oil-in-water emulsion, liposome, aluminum hydroxide gel, and silica adjuvant. Freund's complete adjuvant is widely used and can be suitably used in the present invention. For example, during the first immunization (sensitization), it is preferable to use Freund's complete adjuvant as the adjuvant.

  In the immunization, booster immunization is performed when a predetermined period has elapsed after the first immunization (sensitization). In this additional immunization, it is preferable to add various adjuvants to a solution containing an effective amount of the modified carrier protein. For example, it is preferable to use Freund's complete adjuvant as the adjuvant at the time of booster immunization, but a considerable effect can be obtained by using Freund's incomplete adjuvant.

  After the first immunization operation (sensitization), the booster immunization to be performed is preferably performed a plurality of times. It is desirable to perform booster immunization when the antibody concentration in the blood shows a maximum due to the immune response to the previous immunization operation (sensitization) and the antibody concentration is decreasing. The number of days until the antibody concentration in the blood reaches the maximum after the last immunization (sensitization) usually depends on the metabolic rate in the body of the immunogen used. Thus, the booster interval depends on the type of immunogen used, the type of immunized animal of interest, and its health condition. When a small animal such as a mouse is used as an immunized animal, a form in which booster immunization is performed after the first immunization (sensitization), for example, 2 weeks, 4 weeks, 6 weeks, or 8 weeks can be selected.

  Scientifically, the type of mammal to be immunized is not limited, but is selected from mammals other than humans from an ethical viewpoint. When creating hybridoma cells that produce monoclonal antibodies, mice, rats, goats, and the like can be selected as mammals other than humans that can be used for mammals to be immunized.

  As a mammal to be immunized, a mammal that already holds an antibody showing cross-reactivity with the modified carrier protein is not preferable. That is, it is generally preferable that the mammal to be immunized is an individual who has no acquired immunity. In view of the above requirements, it is preferable to use mammals grown after birth in an environment that is essentially free from exposure to various immunogenic substances. Alternatively, it is preferable to use a mammal having a short period until it grows to the extent that it can be subjected to immunization after giving birth. In consideration of these conditions, it is more preferable to use small pedigree mammals used for medical research. Specifically, mice, rats, rabbits and the like that are used for the creation of various novel antibodies are preferable, and in particular, mice or rats, and more preferably mice are used.

  Prior to immunization, the non-human mammal used as an immunized animal is used to produce the modified carrier protein, the immunogenicity of the carrier protein itself, and a specific antibody against the carrier protein It is desirable to investigate in advance the immunization conditions for which is created. A method in which a low molecular weight organic compound of interest is bound to the above carrier protein and the resulting modified carrier protein is used as an immunogen (Non-patent Document 2: Chemistry Letters, Vol. 35, No. 10, p. 1126-1127 (2006)), regarding the various carrier proteins already used, the above matters have already been described for mice, rats, rabbits, etc., which are used for the creation of various novel antibodies. It has been investigated and reports are available.

  In addition, a method of binding a target low molecular weight organic compound on the carrier protein and using the resulting modified carrier protein as an immunogen (Non-patent Document 2: Chemistry Letters, Vol. 35, No. 10, p.1126-1127 (2006)), referring to the successful examples already reported, the immunization of modified carrier proteins for mice, rats, rabbits, etc., which are used for the creation of various new antibodies It is also possible to estimate the effective amount with considerable accuracy.

  It is desirable to select the procedure of immunization for mice, rats, rabbits, etc., which are used to create various novel antibodies, in accordance with the procedures used in the successful examples already reported.

  When a mouse or rat is selected as the mammal to be immunized, the age at which the first immunization operation (sensitization) is performed is selected in consideration of the number of subsequent immunizations and the interval thereof. Specifically, it is necessary to verify that an antibody specific for the immunogen is present in the blood of the immunized animal after completion of multiple boosters. Therefore, it is preferable to select an age at which the first immunization operation (sensitization) is performed so that the immunized animal does not reach an age at which the ability to produce antibodies decreases at the time of completing multiple boosters. . When the period until the end of multiple boosts after the first immunization (sensitization) is selected to be about 8 weeks, in mice or rats, the age at which the first immunization (sensitization) is performed is: It is preferable to select the range of 10 to 15 weeks of age, and it is usually more preferable to select the range of about 12 weeks of age. It is known that mice or rats have the ability to produce sufficient antibodies when they reach about 12 weeks of age, and the ability to create new antibodies is the highest.

  The dicarboxylic acid compound having the structure represented by the formula (II) is a known compound, and its synthesis method has already been reported in the literature (Non-patent Document 3: Organic & Biomolecular Chemistry Vol. 4, p.4431- 4436 (2006)). Since the dicarboxylic acid compound having the structure represented by the formula (II) is solid at room temperature, the reaction for binding the compound on the carrier protein is performed using a reaction solvent capable of dissolving the compound. There is a need. On the other hand, the carrier protein used may undergo denaturation depending on the type of solvent. Therefore, it is necessary to select a reaction solvent capable of dissolving the compound without causing denaturation of the carrier protein used.

Conventionally, dicarboxylic acids having a structure represented by the above formula (II) from various reaction solvents used in preparing modified carrier protein by binding a target low molecular weight organic compound on carrier protein. It is preferable to select a reaction solvent capable of dissolving the acid compound. In fact, when the selection of the reaction solvent was advanced, dimethyl sulfone (DMSO: (CH ₃ ) ₂ SO) and a borate buffer were selected as particularly preferable reaction solvents.

Dimethylsulfone (DMSO: (CH ₃ ) ₂ SO) is a non-aqueous solvent, but can be dissolved without denaturing the carrier protein, and is equivalent to a dicarboxylic acid compound having the structure shown in formula (II) It is a solvent that can be dissolved at a high concentration.

  Further, the boric acid buffer solution has a pH range in which the buffering action can be exerted in the range of 6.8 to 9.2, and is a solvent capable of dissolving the dicarboxylic acid compound having the structure represented by the above formula (II). As the system, it is usually preferable to select a composition in which the pH is selected in the range of 8.2 to 8.7, particularly a composition in which the pH can be adjusted to around 8.5.

Reactive functional groups present on the surface of the carrier protein using a carboxyl group (—COOH), which is a reactive functional group present in the molecule of the dicarboxylic acid compound having the structure represented by the formula (II), In the case of forming an amide bond (—CO—NH—) with an amino group (—NH ₂ ), it is preferable to use an amide bond forming method using carbodiimide. In amide bond formation using carbodiimide, N, N′-dicyclohexylcarbodiimide (DCC), N, N′-diisopropylcarbodiimide (DIC), N- [3- (dimethylamino) propyl] -N ′ is used as a binder carbodiimide. -Ethylcarbodiimide (EDC), N- [3- (dimethylamino) propyl] -N'-ethylcarbodiimide hydrochloride (EDAC), etc. can be utilized. The amount of the binder carbodiimide is preferably selected in the range of 5 to 20 molecules per molecule of the dicarboxylic acid compound having the structure represented by the formula (II).

On the other hand, the amount of the dicarboxylic acid compound having the structure represented by the formula (II) is determined based on the total number of amino groups (—NH ₂ ) exposed on the surface of the carrier protein. At that time, when the total number of amino groups (—NH ₂ ) exposed on the surface of one molecule of the carrier protein is N, the amount of the dicarboxylic acid compound having the structure represented by the formula (II) is used. It is preferable to select in the range of N × 3 molecules to N × 10 molecules per protein molecule. As a result, a modified carrier protein is prepared in which a dicarboxylic acid compound having a structure represented by the formula (II) is bound in a range of 1/2 × N molecule to N molecule per molecule of the carrier protein. It is desirable to do.

  In the present invention, a modified carrier protein obtained by binding a target low molecular weight organic compound on a carrier protein is used as an immunogen to be used for the above-described immunization. Among the multiple types of antibodies that are newly created by immunization and specific for the modified carrier protein, it is also high against the low molecular weight organic compound itself that is not bound to the carrier protein used. First, it is verified that there is actually an antibody showing reactivity.

  After completion of the above-mentioned last round of booster immunization, when a significant increase in the blood concentration of an antibody specific for the modified carrier protein used as an immunogen is found, blood is collected from the immunized animal and collected. Antiserum is prepared from the collected blood. Among the multiple antibodies specific to the modified carrier protein contained in the antiserum, high reactivity with the low molecular weight organic compound itself that is not bound to the carrier protein used It is verified that an antibody showing is actually present.

  That is, the presence or absence of a polyclonal antibody having the low molecular weight organic compound itself as an antigenic determinant is verified. An enzyme immunoassay (ELISA method) is preferably used as a means for verifying the presence of an antibody that specifically binds to a specific antigenic determinant in antisera containing multiple types of antibodies. The The enzyme immunoassay (ELISA method) uses a specific reactivity of an antibody to a specific antigenic determinant, and therefore has high selectivity, particularly for a specific antigenic determinant contained in the antiserum. When the antibody concentration is unknown, the antibody titer can be easily evaluated.

  In the present invention, in order to detect an antibody that is not bound to the carrier protein used and shows high reactivity with the low molecular weight organic compound itself, it is used in an enzyme immunoassay (ELISA method). As an antigen, another kind of modified carrier protein in which the low molecular weight organic compound is bound to the surface of another kind of carrier protein is used. Of course, it is necessary that the different type of carrier protein itself does not react with a plurality of types of antibodies specific to the modified carrier protein.

  The carrier protein used for the preparation of the immunogen and the other kind of carrier protein used for the preparation of the antigen used in the enzyme immunoassay (ELISA method) are preferably used for the preparation of the immunogen. It is preferable to select a combination of two different carrier proteins from the group of carrier proteins.

  In the carrier protein combination, antigenic determinants of the carrier protein itself are usually different, and multiple types of antibodies specific to the modified carrier protein react with the different carrier protein itself. The possibility of showing sex can be eliminated. Further, in the combination of the two different carrier proteins described above, it is very unlikely that the local structure (partial amino acid sequence) of the site capable of binding the low molecular weight organic compound substantially matches. Therefore, in the above-mentioned combination, another type of modified carrier protein in which the low molecular weight organic compound is bound to the surface of another type of carrier protein among a plurality of types of antibodies specific to the modified carrier protein of the immunogen. An antibody that exhibits binding ability to can be regarded as an antibody that binds to the low molecular weight organic compound itself.

  In particular, bovine serum albumin, which is widely used as a blocking protein, is selected as another type of carrier protein used for the preparation of the antigen used in the enzyme immunoassay (ELISA method), while the production of the immunogen It is more preferable to select a general-purpose carrier protein other than bovine serum albumin, such as Keyhole Limpet Hemocyanin, as the carrier protein used in the above. When bovine serum albumin is selected as another type of carrier protein used for the production of a modified carrier / protein type antigen used in the enzyme immunoassay (ELISA method), the modified carrier / protein type antigen The phenomenon of non-selective binding of antibody molecules to the bovine serum albumin moiety is also eliminated. Further, the modified carrier protein type antigen using bovine serum albumin as a carrier protein can be immobilized on an ELISA plate at a high density.

  After verifying that the obtained antiserum contains an antibody having reactivity to the low molecular weight organic compound itself, which is used to produce an immunogen, the reactivity to the low molecular weight organic compound itself Whether or not a polyclonal antibody having a cross-reactivity with a target peroxide derivative-type explosive, for example, trimeric acetone peroxide (TATP) itself having a structure represented by formula (I), Verify that.

  In the present invention, the verification of the cross-reactivity of the antibody preferably uses a competitive reaction of the two antigens with the antibody.

  The target peroxide derivative-type explosive, for example, trimer acetone (TATP) itself having the structure shown in formula (I), is a low molecular weight organic compound, and is capable of antigen-antibody reaction with an antibody. In doing so, the binding is believed to be achieved by one of the complementarity determining sites of the antibody molecule. In addition, when a low molecular weight organic compound having a similar structure used for the production of a modified carrier / protein used for the above-described immunization is also subjected to an antigen-antibody reaction with an antibody, its binding is caused by an antibody molecule. It is thought that this is achieved by one of the complementarity determining sites.

  Therefore, when an antibody has cross-reactivity, the complementarity determining site of the antibody molecule involved in binding to a low molecular weight organic compound having a similar structure, which is used to produce a modified carrier protein of an immunogen And the target peroxide derivative-type explosive, for example, the complementarity determining site of the antibody molecule involved in the binding of the trimeric acetone peroxide (TATP) itself shown in formula (I). Very likely. In that case, when the target peroxide derivative-type explosive, for example, the trimeric acetone peroxide (TATP) shown in the formula (I) binds to the complementarity determining site of the antibody molecule, the immunogen is modified. It inhibits the binding of low molecular weight organic compounds having a similar structure that are used in the production of carrier proteins. By utilizing this phenomenon of competitive inhibition, it is possible to verify whether or not a specific complementarity determining site of the antibody molecule exhibits cross-reactivity.

  Specifically, the low molecular weight compound having the similar structure used for the verification of the reactivity to the low molecular weight organic compound itself having a similar structure, which is used for the production of the modified carrier / protein of the above immunogen. Another type of modified carrier protein having an organic compound bound to the surface of another type of carrier protein is immobilized on an ELISA plate. On the other hand, the desired peroxide derivative-type explosive, for example, trimer-type acetone peroxide (TATP) represented by the formula (I) is added to the polyclonal solution in the reaction solution for antigen-antibody reaction in the ELISA method. Dissolve with antiserum containing antibody.

  When the above competitive reaction proceeds, the cross-reactive antibody present in the reaction solution is a target peroxide derivative-type explosive, for example, a trimeric acetone peroxide represented by the formula (I) As a result of the antigen-antibody reaction with (TATP), the amount of antibody molecules immobilized is reduced through the antigen-antibody reaction with the modified carrier / protein antigen immobilized on the plate. Due to this competitive reaction, the decrease in the amount of the immobilized antibody through the antigen-antibody reaction with the modified carrier protein type antigen immobilized on the plate is determined by enzyme immunoassay (ELISA method). Detect by applying.

  By using this technique, the polyclonal antibody against the low molecular weight organic compound itself having a similar structure used in the production of the modified carrier protein of the above-mentioned immunogen contained in the antiserum, It is possible to verify that a peroxide derivative-type explosive, such as, for example, a trimer-type acetone peroxide (TATP) represented by the formula (I), and an antibody showing cross-reactivity are included.

  In other words, the antiserum verified as described above is a polyclonal antibody having a binding ability to a target peroxide derivative-type explosive, for example, a trimeric acetone peroxide (TATP) represented by the formula (I). Is included.

  It has been verified that an antibody exhibiting cross-reactivity with the target peroxide derivative-type explosive, for example, trimeric acetone peroxide (TATP) represented by the formula (I) has been produced. An animal antibody-producing cell group is collected, and this antibody-producing cell group is fused with a cell of a cell line derived from myeloma to produce a group of hybridoma cells.

  Usually, a spleen cell group is prepared by removing the spleen from the immunized animal that has been verified as described above. This spleen cell group and myeloma-derived cell line cells are fused to produce a group of hybridoma cells.

  The myeloma-derived cell line used for the above-mentioned cell fusion needs to be compatible with the spleen cell derived from the immunized animal to be fused. The proliferation ability of hybridoma cells created by cell fusion depends on the myeloma-derived cell line used for cell fusion, and it is preferable to use a myeloma-derived cell line with excellent proliferation ability. .

  For example, when a mouse is selected as the immunized animal, the myeloma-derived cell line used for cell fusion is a mouse myeloma-derived cell line such as P3X63 Ag8.653, P3X63Ag8U, Sp2 / O Ag14, FO.1, S194 / 5. XX0 BU. 1 or the like is preferably used. In particular, the use of the cell line P3X63Ag8U has high proliferation ability of the hybridoma cells to be created, and the antibody molecules produced by the hybridoma cells are all antibodies that have been properly assembled, and antibodies that have not been assembled yet More preferred because it does not contain molecular fragments.

  For example, when a rat is selected as the immunized animal, the myeloma-derived cell line used for cell fusion is the rat myeloma-derived cell line 210, RCY3. Ag1.2.3, YB2 / 0, etc. are mentioned.

  Examples of the cell fusion technique for creating the hybridoma cell include a polyethylene glycol method, a method using Sendai virus, and a method using an electric current. The polyethylene glycol method is more suitable for the present invention because it has low cytotoxicity, is easy to fuse, and particularly has high reproducibility. That is, in the present invention, among a group of hybridoma cells producing a specific monoclonal antibody against a modified carrier protein of an immunogen, a target peroxide derivative-type explosive, for example, three of the formula (I) It is necessary to select hybridoma cells that produce monoclonal antibodies having cross-reactivity to monomeric acetone peroxide (TATP). The frequency of the hybridoma cells that produce the cross-reactive monoclonal antibody among the group of hybridoma cells created is never high, so the number of cell lines of the group of hybridoma cells to be screened (number of populations) Need to be larger. Therefore, it is preferable to select a cell fusion method with higher reproducibility, and the polyethylene glycol method meets the above-mentioned requirements.

  The created group of hybridoma cells are dispersed, dispensed into a microplate, and grown under known culture conditions appropriately selected according to the utilized myeloma cell line. For a group of hybridoma cell lines established by the above culture, the monoclonal antibody produced by each hybridoma cell line is the target peroxide derivative-type explosive, for example, three of the formula (I) It is verified whether the antibody is cross-reactive with monomeric acetone (TATP).

  For a group of hybridoma cell lines established by culture, the culture supernatant of each hybridoma cell line is collected. The culture supernatant of each hybridoma cell line contains a monoclonal antibody produced by the cell line.

  Monoclonal antibodies contained in the culture supernatant of each hybridoma cell line have reactivity to low molecular weight organic compounds themselves with similar structures used for the production of modified carrier proteins used for immunization. First, it is verified whether it has or not.

  The verification is based on an enzyme immunoassay (ELISA method) in which a low molecular weight organic compound having a similar structure is bound to the surface of another type of carrier protein and another type of modified carrier protein is used as an antigen. Verification methods can be used. The specific measurement method is in principle the same as the verification regarding the reactivity of the polyclonal antibody contained in the antiserum.

  By this verification, a monoclonal antibody having reactivity to a low molecular weight organic compound itself having a similar structure, used for production of a modified carrier protein used for immunization, from a group of hybridoma cell lines. A cell line of hybridoma cells that produces is selected. With respect to the group of cell lines of hybridoma cells selected by this primary screening, the monoclonal antibody produced by the cell line of hybridoma cells is the target peroxide derivative-type explosive, that is, three of the formula (I) It is verified whether or not the antibody exhibits cross-reactivity to monomeric acetone (TATP).

  The verification regarding the cross-reactivity can be performed by applying the same method in principle as the verification regarding the cross-reactivity of the polyclonal antibody contained in the antiserum.

  The target peroxide derivative type explosive, that is, the target peroxide derivative is verified by the cross-reactivity (secondary screening) with respect to the trimeric acetone peroxide (TATP) represented by the formula (I) A hybridoma cell line is produced that produces a monoclonal antibody that exhibits cross-reactivity to the type of explosive, ie, the trimeric acetone peroxide (TATP) of formula (I). The type of monoclonal antibody selected depends on the antibody type specificity exhibited by the anti-Ig antibody used in the enzyme immunoassay (ELISA method).

  Binding to the target peroxide derivative-type explosive, for example, trimeric acetone peroxide (TATP) represented by the formula (I) using the hybridoma cell line selected by this secondary screening Monoclonal antibodies having the ability can be produced.

  The selected hybridoma cell line can be cultured in vitro, the culture supernatant can be collected, and the contained monoclonal antibody can be purified. Further, when the selected hybridoma cell line is inoculated into the abdominal cavity of a mammal other than a human used for immunization, it grows in the abdominal cavity and accumulates the monoclonal antibody produced in the ascites. Thereafter, the ascites can be collected and the contained monoclonal antibody can be purified.

  For purification of monoclonal antibodies contained in ascites or culture supernatant, for example, DEAE anion exchange chromatography, affinity chromatography, ammonium sulfate fractionation method, PEG fractionation method, ethanol fractionation method, etc. Purification to a purity of Desirable purity is 95% or more, more preferably 98% or more. For example, a target peroxide derivative-type explosive, for example, a monoclonal antibody having binding ability to trimeric acetone peroxide (TATP) represented by the formula (I) is immobilized on the peroxide derivative-type explosive. In the case of use in the purification, it is desirable to carry out purification to the above purity.

(Trust number)
In addition,
As a hybridoma cell producing a monoclonal antibody having a binding ability to the trimeric acetone peroxide (TATP) represented by the formula (I) used in the present invention,
Hybridoma cell line: NECP-C57Z 3B-7E is based on the Budapest Treaty. National Institute of Advanced Industrial Science and Technology, Patent Biological Deposit Center In addition, an international deposit (May 12, 2009) has been made.

  The hybridoma cell line is a hybridoma cell line created and selected by the procedure disclosed in the first embodiment to be described later. The hybridoma cell line: NECP-C57Z 3B-7E is a hybridoma cell line that produces the monoclonal antibody mAb-T003 described below.

  In the present invention, it is preferable to prepare a wiping treatment material having the following constitution by using a monoclonal antibody having binding ability to trimeric acetone peroxide (TATP) represented by the formula (I).

  The explosive wiping treatment material according to the first embodiment of the present invention includes a liquid-absorbing material having a function of sucking the liquid in which the explosive is dissolved, for example, the water-absorbing material 1 and the trimmer shown in the formula (I). It has a configuration in which a monoclonal antibody 2 having a binding ability to body-type acetone peroxide (TATP) is held. An example of the explosive wiping treatment material according to the first embodiment of the present invention will be described in detail with reference to FIG.

  An example of the explosive wiping treatment material according to the first embodiment of the present invention illustrated in FIG. 2 is based on a liquid-absorbing material having a function of sucking a liquid in which the explosive is dissolved, for example, a water-absorbing material 1. As an agent, the monoclonal antibody 2 is immobilized on the base by using a polyvinyl alcohol 10 coating layer.

  The liquid-absorbing material used is a material having a liquid-absorbing property with respect to a solvent capable of dissolving acetone peroxide of the formula (I). Specifically, the liquid absorbing property of the liquid absorbing material indicated by the liquid absorbing material is maintained in the state in which the solvent enters the gap space existing in the water absorbing material 1 by capillary action. Is achieved. Therefore, the liquid-absorbing material used is a material having a shape that holds a minute gap space at a high density therein, for example, a sponge shape, a cotton shape, and a non-woven fabric shape. The substance constituting the liquid-absorbing material is one in which the solvent exhibits good wettability. For example, when an aqueous solvent is used when dissolving acetone peroxide of the formula (I), a water-absorbing material is suitably used as the liquid-absorbing material. In order for the solvent to exhibit good wettability, the substance constituting the liquid-absorbing material preferably has a high affinity for the solvent. For example, when an aqueous solvent is used, it is preferable that the substance constituting the liquid-absorbing material has a high affinity for water. Specifically, when an aqueous solvent is used, it is preferable to select polyurethane, cellulose, or a cellulose derivative as a substance constituting the liquid absorbing material. For example, when an aqueous solvent is used, a sponge-like polyurethane, a cotton-like or nonwoven-like material composed of fibers mainly composed of cellulose or a cellulose derivative is preferably used as the liquid-absorbing material.

  The size of the gap space present in the material showing the above-mentioned sponge shape, cotton shape, and nonwoven fabric shape is selected to a size that allows the solvent to enter the gap space by capillarity and maintain the infiltrated state. The Actually, the size of the gap space is selected in consideration of the wettability of the substance constituting the liquid-absorbing material with the solvent, that is, the affinity for the solvent.

  The total volume occupied by the interstitial spaces present in the liquid-absorbing material needs to be larger than the amount of the solution in which acetone peroxide of formula (I) is to be absorbed. In the liquid-absorbing material, the ratio of the volume occupied by the gap space, that is, the porosity is preferably selected in the range of 50% to 90% or 60% to 80% in the dry state.

  When the solution is absorbed into the gap space, the bulk volume of the entire liquid-absorbing material may increase and swell compared with the dry state. In the case of a swellable liquid-absorbing material, the porosity in the dry state is lower than the above range, but the volume ratio occupied by the infiltrated solution after swelling is preferably in the above range.

  When the explosive wiping treatment material according to the first embodiment of the present invention is used for the work of dissolving and wiping off the solid powder of acetone peroxide of formula (I) with an aqueous solvent, the liquid absorbing material As above, it is preferable to select a material having a sponge shape, a cotton shape, or a non-woven fabric shape, which is composed of a substance having a high affinity for water.

  An example of a procedure for immobilizing a monoclonal antibody having binding ability to acetone peroxide of formula (I) in the liquid absorbing material is shown below.

  The liquid-absorbing material is immersed in a solution in which a monoclonal antibody capable of binding to acetone peroxide of the formula (I) to be fixed is dissolved. By this immersion treatment, the entire liquid-absorbing material is infiltrated with the monoclonal antibody solution.

  The infiltrated absorbent material taken out of the monoclonal antibody solution is dried. In the drying process, evaporation of the solvent contained in the solution proceeds from the upper surface of the liquid-absorbing material. The monoclonal antibody contained in the solution is non-selectively adsorbed on the surface of the structure composed of a solvophilic substance constituting the liquid-absorbing material. Accompanying the drying, the monoclonal antibody is fixed on the surface of the structure composed of the solvophilic substance.

  For example, in a sponge sheet-like liquid-absorbing material, when the evaporation of the solvent contained in the solution proceeds, the concentration of the monoclonal antibody solution proceeds. In the partially dried state, the concentrated solution is in a state in which only the region on the lower surface side of the liquid-absorbing material is infiltrated due to its own weight. The adsorption density of the monoclonal antibody that non-selectively adsorbs on the surface of the structure composed of the solvophilic substance constituting the liquid-absorbing material is proportional to the concentration of the monoclonal antibody in the solution. Therefore, due to the concentration of the solution accompanying the progress of drying, the adsorption density of the monoclonal antibody increases from the upper surface to the lower surface of the liquid absorbing material. When the thickness from the lower surface is d, the adsorption density of the monoclonal antibody: Cab (d) shows a distribution that approximately satisfies the relationship of Cab (d) ∝1 / d. That is, as compared with the upper surface side of the liquid-absorbing material, the monoclonal antibody is fixed at a relatively high density on the lower surface side.

  The monoclonal antibody solution used in the above-described fixing process of the monoclonal antibody is usually prepared using an aqueous solvent. Specifically, it is prepared using a buffer compatible with the monoclonal antibody. The concentration of the monoclonal antibody dissolved in the solution is selected according to the amount of acetone peroxide solid powder of formula (I) to be wiped off. Specifically, when the liquid absorbing material is infiltrated, the amount (molar amount) of the monoclonal antibody present in the liquid volume of the monoclonal antibody solution that penetrates into the liquid absorbing material is wiped off. It is selected according to the amount (molar amount) of solid powder of acetone peroxide of I). Usually, the amount (molar amount) of the monoclonal antibody present in the liquid amount of the monoclonal antibody solution is equal to or larger than the amount (molar amount) of the solid powder of acetone peroxide of formula (I) to be wiped off. Preferably, the concentration of monoclonal antibody that is dissolved in the solution is selected. However, when the evaporation of the solvent contained in the solution proceeds and the concentration proceeds, the concentration of the monoclonal antibody contained exceeds the saturation concentration even when the concentration of the monoclonal antibody increased to about 4 times the initial concentration. It is desirable that there is no range. The initial concentration of the monoclonal antibody solution used for the immersion treatment is usually selected in the range of 10 mg / ml to 20 mg / ml, preferably in the range of 12 mg / ml to 18 mg / ml.

  In order to immobilize the monoclonal antibody fixed on the surface of the structure made of the solvophilic substance, for example, a coating layer made of polyvinyl alcohol is formed on the surface of the structure made of the solvophilic substance. Specifically, the liquid-absorbing material after the drying treatment is infiltrated with a solution in which polyvinyl alcohol is dissolved. Even in the process of drying the infiltrated liquid-absorbing material, the solvent contained in the polyvinyl alcohol solution evaporates mainly from the upper surface of the liquid-absorbing material. In the infiltrated state, the polyvinyl alcohol contained in the solution constitutes an adsorbing layer that covers the entire surface of the structure composed of the solvophilic substance. As the solvent evaporates, the polyvinyl alcohol The adsorption layer is converted into a coating layer made of polyvinyl alcohol.

  For example, in a sponge sheet-like absorbent material, when the evaporation of the solvent contained in the polyvinyl alcohol solution proceeds, the concentration of the polyvinyl alcohol solution proceeds. In the partially dried state, the concentrated solution is in a state in which only the region on the lower surface side of the liquid-absorbing material is infiltrated due to its own weight. The adsorption density of polyvinyl alcohol adsorbed on the surface of the structure composed of a solvophilic substance constituting the liquid absorbing material is proportional to the concentration of the polyvinyl alcohol solution. Therefore, due to the concentration of the solution accompanying the progress of drying, the adsorption density of the polyvinyl alcohol increases from the upper surface to the lower surface of the liquid absorbing material. When the thickness from the lower surface is d, the adsorption density of polyvinyl alcohol: Cpv (d) shows a distribution that approximately satisfies the relationship of Cpv (d) ∝1 / d. That is, as compared with the upper surface side of the liquid-absorbing material, the lower surface side is relatively coated with polyvinyl alcohol at a higher density.

  The polyvinyl alcohol solution used in the process of forming the polyvinyl alcohol coating layer is usually prepared using an aqueous solvent. Specifically, it is prepared using an aqueous solvent having good wettability with respect to the lyophilic substance constituting the liquid-absorbing material. It is preferable to use an aqueous solvent that is already fixed and does not exhibit a function of promoting the release of the monoclonal antibody. The concentration of polyvinyl alcohol dissolved in the solution is selected according to the total inner surface area of the gap space of the liquid-absorbing material that forms the coating layer. Specifically, when the liquid absorbing material is infiltrated, the amount of polyvinyl alcohol present in the liquid volume of the polyvinyl alcohol solution that penetrates into the liquid absorbing material is the amount of the liquid absorbing material that should form the coating layer. It is selected according to the total inner surface area of the gap space and the film thickness of the coating layer to be formed. Usually, the average film thickness of the polyvinyl alcohol coating layer to be formed is selected in the range of 0.5 μm to 5 μm. In the case of the above average film thickness range, the initial concentration of the polyvinyl alcohol solution used for the infiltration treatment is usually in the range of 0.5 w / v% to 3 w / v%, preferably 1 w / v%. It is preferable to select in the range of ˜3 w / v%.

  Since the solid powder of acetone peroxide of the formula (I) is used for the work of dissolving and wiping off with an aqueous solvent, the formed coating layer exhibits high hydrophilicity, and the coating layer is water-soluble. Since it is necessary to have permeability, a coating layer made of polyvinyl alcohol can be suitably used. The formed coating layer exhibits high hydrophilicity, and the coating layer can be formed using a material other than polyvinyl alcohol as long as it has water permeability. In addition to polyvinyl alcohol, for example, a form in which a coating layer is formed using a hydrophilic polymer material such as gelatin can also be employed.

  The explosive wiping treatment material according to the second embodiment of the present invention includes a liquid absorbing material having a function of sucking a liquid in which the explosive is dissolved, for example, a water absorbing material 3 and a trimer shown in the formula (I). The monoclonal antibody 2 having the binding ability to the body form of acetone peroxide (TATP) is immobilized via a linker 9, for example, a protein. An example of the explosive wiping treatment material according to the second embodiment of the present invention will be described in detail with reference to FIG.

  In an example of the explosive wiping treatment material according to the second embodiment of the present invention illustrated in FIG. 3, a liquid absorbing material having a function of absorbing an aqueous solution in which the explosive is dissolved, for example, a water absorbing material 3. As a base, a linker 9 having a high binding ability to the monoclonal antibody 2 is immobilized in advance on the base, and the monoclonal antibody 2 is immobilized through the binding between the linker 9 and the monoclonal antibody 2. Is going on. Furthermore, dissociation of the immobilized monoclonal antibody 2 is prevented by providing a coating layer of polyvinyl alcohol 10.

  The liquid-absorbing material used is a water-absorbing material having a liquid-absorbing property with respect to an aqueous solution in which acetone peroxide of formula (I) is dissolved. Specifically, the water absorption property of the water-absorbing material indicated by the water-absorbing material is that the water-based solvent enters the gap space existing in the water-absorbing material 3 by capillary action and is maintained in the infiltrated state. To be achieved. Therefore, the water-absorbing material used is a water-absorbing material having a shape that holds a minute gap space at a high density therein. Moreover, as for the substance which comprises this water absorbing material, an aqueous solvent shows the favorable wettability. The substance constituting the liquid-absorbing material preferably has, for example, high affinity for water contained in the aqueous solvent and excellent ability to retain the aqueous solvent that enters the gap space. Specifically, Sepurafilm® of Genzyme, which is a film-like water-absorbing material containing hyaluronic acid having excellent water retention and carboxymethylcellulose as components, can be suitably used.

  When these water-absorbing materials having water retention are infiltrated with an aqueous solvent, the bulk volume of the entire water-absorbing material is slightly increased and swollen as compared with the dry state. It is preferable that the volume ratio of the infiltrated aqueous solvent after the swelling is usually in the range of 5% to 35% and in the range of 10% to 20%.

  On the other hand, the water-absorbing material 3 has a carboxyl group as a hydrophilic group present on the sugar chain skeleton of the constituent components of the fiber, and a linker 9 such as a monoclonal antibody 2 is utilized using the carboxyl group. In contrast, protein molecules having high binding ability are immobilized.

  Specifically, the water-absorbing material 3 has a carboxyl group activated, and the linker protein molecule is converted into the linker protein molecule by reacting the activated carboxyl group with the linker protein molecule. It is immobilized covalently on the surface of the fiber material constituting the water absorbing material 3. The activation of the carboxyl group can be performed, for example, by a method using carbodiimide and N-hydroxysuccinimide. The method using carbodiimide and N-hydroxysuccinimide can easily and reliably activate the carboxyl group exposed on the surface of the gap space of the water-absorbing material. In the second embodiment of the present invention, Are preferably used.

  For example, in the method using carbodiimide and N-hydroxysuccinimide, N, N′-dicyclohexylcarbodiimide (DCC), N, N′-diisopropylcarbodiimide (DIC), N- [3- (dimethylamino) propyl is used as the carbodiimide. ] -N'-ethylcarbodiimide (EDC), N- [3- (dimethylamino) propyl] -N'-ethylcarbodiimide hydrochloride (EDAC), and the like can be used.

  Protein A and protein G can be suitably used as protein molecules having high binding ability to the monoclonal antibody 2 that can be used as the linker 9. Protein A and protein G are widely used for immobilization using the carboxyl group subjected to the above activation treatment, and also have high binding ability to antibody molecules and are inexpensive. The linker used in the second embodiment is suitable. Of course, proteins other than protein A and protein G can also be used as the linker 9 as long as the protein can be immobilized using a carboxyl group subjected to activation treatment and has a high binding ability to antibody molecules.

  The concentration of the linker 9 contained in the solution in which the linker 9 is dissolved used for the immobilization reaction is selected according to the amount of the monoclonal antibody 2 to be finally immobilized on the water absorbing material 3. That is, the amount (molar amount) of the linker 9 immobilized in advance on the water absorbing material 3 is equal to or larger than the amount (molar amount) of the monoclonal antibody 2 to be finally immobilized on the water absorbing material 3. The concentration of linker 9 contained in the solution is selected. Specifically, the concentration of the linker 9 contained in the solution used for the immobilization reaction, for example, the concentration of protein A or protein G is usually in the range of 0.5 mg / ml to 10 mg / ml, preferably Is preferably selected in the range of 1 mg / ml to 5 mg / ml.

  The water-absorbing material 3 that has completed the immobilization of the linker 9 is immersed in an aqueous solution containing the monoclonal antibody 2. By performing this immersion treatment, the monoclonal antibody 2 is bound to the linker 9 fixed to the water absorbing material 3. Thereafter, even after the aqueous solution containing the monoclonal antibody 2 is washed and removed, the monoclonal antibody 2 is held in a state of being bonded to the linker 9. After the washing, the water absorbing material 3 is dried. Even in this state, the monoclonal antibody 2 is in a state of being bound to the linker 9, and thereafter, when the aqueous solvent is absorbed, the dissociation of the monoclonal antibody 2 proceeds only slightly.

  When binding to the linker 9, the concentration of the monoclonal antibody 2 contained in the aqueous solution is usually selected in the range of 10 mg / ml to 20 mg / ml, preferably in the range of 12 mg / ml to 18 mg / ml. preferable.

  In the example shown in FIG. 3, the dissociation of the monoclonal antibody 2 is further prevented by forming a coating layer of polyvinyl alcohol 10.

  The polyvinyl alcohol solution used in the process of forming the polyvinyl alcohol 10 coating layer is usually prepared using an aqueous solvent. Specifically, it is prepared using an aqueous solvent having good wettability with respect to the hydrophilic substance constituting the water absorbing material 3. The concentration of polyvinyl alcohol dissolved in the solution is selected according to the total inner surface area of the gap space of the water absorbing material 3 that forms the coating layer. Specifically, when the water-absorbing material 3 is infiltrated with an aqueous solution, the amount of polyvinyl alcohol that enters the water-absorbing material 3 and is present in the liquid amount of the polyvinyl alcohol solution should form the coating layer. It is selected according to the total inner surface area of the gap space of the material 3 and the film thickness of the coating layer to be formed. Usually, the average film thickness of the polyvinyl alcohol coating layer to be formed is selected in the range of 0.5 μm to 5 μm. When the average film thickness is in the range, the concentration of the polyvinyl alcohol solution used for the infiltration treatment is usually in the range of 0.5 w / v% to 3 w / v%, preferably 1 w / v% to It is preferable to select in the range of 3 w / v%.

  Since the solid powder of acetone peroxide of the formula (I) is used for the work of dissolving and wiping off with an aqueous solvent, the formed coating layer exhibits high hydrophilicity, and the coating layer is water-soluble. Since it is necessary to have permeability, a coating layer made of polyvinyl alcohol can be suitably used. The formed coating layer exhibits high hydrophilicity, and the coating layer can be formed using a material other than polyvinyl alcohol as long as it has water permeability. In addition to polyvinyl alcohol, for example, a form in which a coating layer is formed using a hydrophilic polymer material such as gelatin can also be employed.

  In the explosive wiping and removing method according to the present invention, acetone peroxide (TATP) of the formula (I) is used by using the explosive wiping treatment material according to the first embodiment or the second embodiment. The solid powder can be wiped off and removed according to the procedure described below.

  That is, a solid powder of acetone peroxide of the formula (I) is dissolved in an appropriate amount of a solvent capable of dissolving the acetone peroxide of the formula (I). By absorbing the solution of acetone peroxide of formula (I) dissolved in the appropriate amount of solvent in the explosive wiping treatment material according to the first embodiment or the second embodiment, A wiping treatment of a solution in which acetone peroxide of formula (I) is dissolved is performed.

  An example of a specific procedure for performing the above series of operations will be described with reference to FIGS.

  In the procedure shown in FIG. 4, as the sheet-like wiping treatment material, the water-absorbing material 4 holding the antibody, which is produced by the procedure described in the first embodiment or the second embodiment, is used. Yes.

  FIG. 4A schematically shows a state in which the solid powder 5 of acetone peroxide (TATP) of the formula (I) remains on the glass substrate 6. The sheet-like wiping treatment material 4 used for wiping and removing is initially in a dry state.

  FIG. 4B schematically illustrates an operation in which an appropriate amount of the solvent 8 used for dissolving the acetone peroxide (TATP) of the formula (I) is dropped with a pipette 7 onto the upper surface of the sheet-like wiping treatment material 4. Indicate. Specifically, the entire sheet-like wiping treatment material 4 is infiltrated with the solvent 8.

  FIG. 4C shows a sheet-like wiping treatment material 4 that has been infiltrated with the solvent 8, acetone peroxide of formula (I) whose lower surface remains on the glass substrate 6 ( (TATP) is schematically shown in a state where it is placed so as to cover the entire solid powder 5. In this state, the solid powder 5 of acetone peroxide (TATP) of the formula (I) in contact with the lower surface of the sheet-like wiping treatment material 4 is infiltrated by the solvent 8 and gradually dissolves. On the lower surface of the sheet-like wiping treatment material 4, the concentration of acetone peroxide (TATP) dissolved in the solvent 8 is initially a saturated concentration. On the other hand, in the solvent 8 on the upper surface side of the sheet-like wiping treatment material 4, the concentration is zero, and in the sheet-like wiping treatment material 4, a concentration gradient of acetone peroxide (TATP) exists. Due to this concentration gradient, the diffusion of dissolved acetone peroxide (TATP) proceeds from the lower surface side of the sheet-like wiping treatment material 4 toward the upper surface side. Accordingly, the solid powder 5 of acetone peroxide (TATP) that was in contact with the lower surface of the sheet-like wiping treatment material 4 is completely dissolved in the solvent 8 and infiltrated with the sheet-like wiping treatment material 4. 8, a state of being dissolved at a substantially uniform concentration is achieved.

  In the sheet-like wiping treatment material 4, a monoclonal antibody having an ability to bind to acetone peroxide of formula (I) is immobilized, and when an antigen-antibody reaction proceeds in the solvent 8, Then, acetone peroxide (TATP) dissolved in the solvent 8 is bound. At the stage where the antigen-antibody reaction has sufficiently progressed in the solvent 8, the amount of the immobilized monoclonal antibody (molar amount) is the amount of the solid powder 5 of acetone peroxide (TATP) of formula (I) (molar amount). In the case where the amount is sufficiently larger than that, acetone peroxide (TATP) dissolved in the solvent 8 reaches a state where substantially all of them are bound to the monoclonal antibody. In other words, the concentration of acetone peroxide (TATP) dissolved in the solvent 8 infiltrated in the sheet-like wiping treatment material 4 reaches a state substantially corresponding to zero.

  When the above state was reached, when the sheet-like wiping treatment material 4 infiltrated with the acetone peroxide solution of formula (I) was removed from the glass substrate 6, it remained on the surface of the glass substrate 6. The solid peroxide 5 of acetone peroxide (TATP) of the formula (I) is in a state of being wiped off. On the surface of the glass substrate 6, an extremely thin liquid film of the solvent 8 may remain in the region that has been in contact with the lower surface of the sheet-like wiping treatment material 4. Even in that case, since the concentration of acetone peroxide (TATP) dissolved in the solvent 8 is substantially equivalent to zero, the remaining extremely thin liquid film of the solvent 8 remains. The amount of acetone peroxide (TATP) contained therein is substantially zero.

  On the other hand, the sheet-like wiping treatment material 4 removed from the glass substrate 6 is in a state infiltrated with the solvent 8, but the concentration of acetone peroxide (TATP) dissolved in the solvent 8 is substantially Therefore, it is in a state corresponding to zero. Therefore, when the sheet-like wiping treatment material 4 used for wiping and removing is incinerated, it can be safely incinerated.

  Even in the procedure shown in FIG. 5, the sheet-like wiping treatment material is produced by using the water-absorbing material 4 holding the antibody, which is produced by the procedure described in the first embodiment or the second embodiment. Yes.

  FIG. 5A schematically shows a state in which the solid powder 5 of acetone peroxide (TATP) of the formula (I) remains on the glass substrate 6. The sheet-like wiping treatment material 4 used for wiping and removing is initially in a dry state.

  FIG. 5B shows an appropriate amount of the solvent 8 used for dissolving the acetone peroxide (TATP) of the formula (I) in an appropriate amount of the acetone peroxide (TATP) solid powder 5 remaining on the glass substrate 6. An operation of dropping with a pipette 7 is schematically shown on the upper surface. Specifically, the whole solid powder 5 of acetone peroxide (TATP) is infiltrated with the solvent 8.

  (C) of FIG. 5 is a sheet-like wiping so that the lower surface of the sheet-like wiping treatment material 4 covers the entire solid powder 5 of acetone peroxide (TATP) in a state infiltrated with the solvent 8. A state in which the treatment material 4 is placed on the glass substrate 6 is schematically shown. In this state, the solid powder 5 of acetone peroxide (TATP) of the formula (I) in contact with the lower surface of the sheet-like wiping treatment material 4 is infiltrated by the solvent 8 and gradually dissolves. The solvent 8 enters from the lower surface of the sheet-like wiping treatment material 4, and the sheet-like wiping treatment material 4 is infiltrated. Initially, the solid powder 5 of acetone peroxide (TATP) is partially dissolved in the dropped solvent 8, but most of the solid powder 5 is not yet dissolved. As a result, the concentration of acetone peroxide (TATP) in the solvent 8 infiltrating the sheet-like wiping treatment material 4 is in a low state.

  Thereafter, dissolution of the solid powder 5 of acetone peroxide (TATP) proceeds on the lower surface side of the sheet-like wiping treatment material 4, and therefore, in this portion, acetone peroxide (TATP) dissolved in the solvent 8 is dissolved. The concentration is a saturated concentration. On the other hand, the concentration is low in the solvent 8 on the upper surface side of the sheet-like wiping treatment material 4, and there is a gradient of acetone peroxide (TATP) concentration in the sheet-like wiping treatment material 4. Due to this concentration gradient, the diffusion of dissolved acetone peroxide (TATP) proceeds from the lower surface side of the sheet-like wiping treatment material 4 toward the upper surface side. Accordingly, the solid powder 5 of acetone peroxide (TATP) that was in contact with the lower surface of the sheet-like wiping treatment material 4 is completely dissolved in the solvent 8 and infiltrated with the sheet-like wiping treatment material 4. 8, a state of being dissolved at a substantially uniform concentration is achieved.

  In the sheet-like wiping treatment material 4, a monoclonal antibody having an ability to bind to acetone peroxide of formula (I) is immobilized, and when an antigen-antibody reaction proceeds in the solvent 8, Then, acetone peroxide (TATP) dissolved in the solvent 8 is bound. At the stage where the antigen-antibody reaction has sufficiently progressed in the solvent 8, the amount of the immobilized monoclonal antibody (molar amount) is the amount of the solid powder 5 of acetone peroxide (TATP) of formula (I) (molar amount). In the case where the amount is sufficiently larger than that, acetone peroxide (TATP) dissolved in the solvent 8 reaches a state where substantially all of them are bound to the monoclonal antibody. In other words, the concentration of acetone peroxide (TATP) dissolved in the solvent 8 infiltrated in the sheet-like wiping treatment material 4 reaches a state substantially corresponding to zero.

  When the above state was reached, when the sheet-like wiping treatment material 4 infiltrated with the acetone peroxide solution of formula (I) was removed from the glass substrate 6, it remained on the surface of the glass substrate 6. The solid peroxide 5 of acetone peroxide (TATP) of the formula (I) is in a state of being wiped off. On the surface of the glass substrate 6, an extremely thin liquid film of the solvent 8 may remain in the region that has been in contact with the lower surface of the sheet-like wiping treatment material 4. Even in that case, since the concentration of acetone peroxide (TATP) dissolved in the solvent 8 is substantially equivalent to zero, the remaining extremely thin liquid film of the solvent 8 remains. The amount of acetone peroxide (TATP) contained therein is substantially zero.

  FIG. 5D schematically shows a state in which acetone peroxide (TATP) is bound to the immobilized monoclonal antibody in the sheet-like wiping treatment material 4 removed from the glass substrate 6.

  On the other hand, the sheet-like wiping treatment material 4 removed from the glass substrate 6 is in a state infiltrated with the solvent 8, but the concentration of acetone peroxide (TATP) dissolved in the solvent 8 is substantially Therefore, it is in a state corresponding to zero. Therefore, when the sheet-like wiping treatment material 4 used for wiping and removing is incinerated, it can be safely incinerated.

  In the wiping and removing step, unnecessary solid friction or impact is not applied to the solid acetone peroxide powder in a dry state, and the solid powder is first infiltrated with a solvent. The solid powder is gradually dissolved in the solvent by keeping it infiltrated with the solvent and allowing to stand at room temperature (for example, 25 ° C.). That is, the solid powder of acetone peroxide (TATP) of formula (I) is sensitive to friction and impact in the dry state, but by infiltrating the solvent, the sensitivity to friction and impact is greatly reduced. I am letting. Further, when the solution is dissolved in a solvent and kept in an infiltrated state and left standing at room temperature (for example, 25 ° C.), the sensitivity to friction and impact is further reduced.

  At that time, the wiping treatment material covering the solid powder of acetone peroxide infiltrated with the solvent is also infiltrated with the solvent, and the base material of the wiping treatment material is in a state of flexibility. Therefore, the above wiping and removing process eliminates the possibility of unnecessary friction and impact on the solid powder of acetone peroxide.

  Therefore, it is necessary to dissolve all the solid powder of acetone peroxide to be removed in a solvent to form a solution. Usually, the amount of solvent used is the amount of solid powder of acetone peroxide to be removed, Based on the solubility in the solvent, the amount of liquid that can achieve complete dissolution is selected.

  The solvent used for dissolving the solid peroxide of acetone peroxide to be removed is selected from solvents that can dissolve the acetone peroxide of the formula (I), and the acetone peroxide of the formula (I) has a high solubility. It is preferable to select from the solvents shown.

  Since the solution of acetone peroxide of formula (I) is in a state of being absorbed by the wiping treatment material, it penetrates into the fine gap space existing in the base material of the wiping treatment material by capillary action. There is a need. Therefore, the solvent itself needs to exhibit sufficient fluidity, and it is usually preferable that the liquid viscosity be low enough to allow entry into a fine gap space by capillary action. In addition, in the solution infiltrated in the fine gap space existing in the base material of the wiping treatment material, the formula (I) existing on the lower surface side in contact with the solid powder of acetone peroxide and on the upper surface side. The diffusion process due to the concentration gradient of acetone peroxide). In order to advance this diffusion process quickly, it is preferable that the solvent viscosity of the solvent itself is low.

  Water is a solvent having a low liquid viscosity and suitable for dissolving the acetone peroxide of the formula (I). Accordingly, water and an aqueous solvent such as a buffer solution used for the antigen-antibody reaction can be used as the solvent used for dissolving the solid powder of acetone peroxide of formula (I).

  In addition, examples of the organic solvent having low liquid viscosity and high solubility of acetone peroxide of the formula (I) include acetonitrile, methanol, ethanol, acetone, ethanolamine, and methyl ethyl ketone. Solvent containing at least one organic solvent selected from the group of organic solvents consisting of acetonitrile, methanol, ethanol, acetone, ethanolamine, methyl ethyl ketone as a solvent used for dissolving solid powder of acetone peroxide of formula (I) Can be suitably used. The solvent containing the organic solvent is usually in the form of a mixture of water and an aqueous solvent such as a buffer used for antigen-antibody reaction and the organic solvent. The content ratio of the organic solvent contained in the mixed solution is generally in the range of 50 v / v% or less, usually in the range of 5 v / v% to 50 v / v%, preferably 5 v / v% to 25 v / v. It is desirable to select in the range of%.

  In the process of dissolving the solid powder of acetone peroxide of formula (I) using the mixed solution, dissolution of acetone peroxide of formula (I) proceeds due to the high affinity exhibited by the organic solvent. When the content ratio of the organic solvent contained in the mixed solution is 50 v / v% or less, for example, 25 v / v% or less, the dissolved acetone peroxide of formula (I) is initially solvated by the organic solvent. However, the ratio of solvated by water molecules is increased by the exchange of organic solvent molecules and water molecules. The antigen-antibody reaction with the monoclonal antibody mainly proceeds with acetone peroxide of formula (I) solvated by water molecules. As the concentration of acetone peroxide of formula (I) solvated by water molecules decreases with the progress of the antigen-antibody reaction, the solvent is dissolved by the water molecules from the state solvated by the organic solvent due to the equilibrium relationship. Conversion to the summed state proceeds. Finally, acetone peroxide of the formula (I) dissolved in the mixed solution is in a state of forming a complex with the monoclonal antibody by the antigen-antibody reaction, and dissolved in the mixed solution. The concentration of acetone peroxide in I) is very low, as determined by the dissociation equilibrium of the monoclonal antibody and the complex.

  On the other hand, when the content ratio of the organic solvent contained in the mixed solution exceeds 50 v / v%, the equilibrium relationship between the state solvated by the organic solvent and the state solvated by the water molecule Therefore, the conversion process from the state solvated by the organic solvent to the state solvated by the water molecule is suppressed. This phenomenon becomes more prominent as the affinity of the organic solvent for acetone peroxide of formula (I) is higher than the affinity of water molecules. In addition, when a plurality of types of organic solvents are added, the equilibrium relationship for transition from a state solvated by a plurality of types of organic solvents to a state solvated by a water molecule is determined by a single organic solvent. Compared with the case where it adds, a branch point transfers to the area | region where the content rate of an organic solvent is relatively low. In other words, there is a competitive relationship between the process of substituting one type of organic solvent with another type of organic solvent and the process of substituting with water molecules, so the transition to a state solvated by water molecules occurs. For this purpose, it is necessary to further increase the content ratio of water molecules. Considering this point, it is more preferable to select the content ratio of the organic solvent contained in the mixed solution in the range of 5 v / v% to 25 v / v%.

  On the other hand, in the method of wiping off a peroxide derivative-type explosive according to the present invention, a monoclonal antibody immobilized in the wiping treatment material and capable of binding to acetone peroxide of formula (I), and a solvent An antibody antigen reaction is performed with acetone peroxide of formula (I) dissolved therein. That is, the solvent used for dissolving the acetone peroxide of formula (I) needs to be a solvent capable of performing the antigen-antibody reaction.

  Of course, the above-mentioned water and an aqueous solvent such as a buffer solution used for the antigen-antibody reaction are solvents that satisfy the above-mentioned requirements.

  When using the above-mentioned solvent containing an organic solvent, it is preferable to use an organic solvent capable of performing the antigen-antibody reaction or a solvent mixture containing an organic solvent. Specifically, the complex of acetone peroxide of the formula (I) and the monoclonal antibody formed by the antigen-antibody reaction in the organic solvent or solvent mixture containing the organic solvent does not easily dissociate. Is preferred.

  In the method of wiping off a peroxide derivative-type explosive according to the present invention, the dissolution process of the solid powder of acetone peroxide of formula (I), the dissolved acetone peroxide of formula (I) and the monoclonal antibody described above After finishing the antigen-antibody reaction process, the wiping treatment material infiltrated with the solution is removed. In the process of removing the wiping treatment material, the concentration of acetone peroxide of the formula (I) dissolved in the solution is substantially zero, but the solid powder of acetone peroxide to be removed is It is preferable to minimize the amount of solution remaining on the surface that was present. Therefore, in the process of removing the wiping treatment material, it is preferable to eliminate the situation where the wiping treatment material is compressed and the infiltrated solution is pushed out.

  Therefore, in the method of wiping and removing a peroxide derivative-type explosive according to the present invention, the wiping treatment material covers the solid surface powder of acetone peroxide of formula (I) to be removed, and the lower surface of the wiping treatment material. It is preferable that there is no gap between the surfaces on which the solid powder of acetone peroxide is present, and the surface is in close contact. That is, instead of the operation of scraping the removal target existing on the surface with a wiping treatment material, such as a general wiping treatment, an operation of covering the removal target with the wiping treatment material and sucking the formed solution is selected. It is preferable.

  The first embodiment and the second embodiment of the method for wiping off a peroxide derivative explosive according to the present invention will be described as an example, and the present invention will be described more specifically.

  Specific examples of the first embodiment and the second embodiment illustrated below are examples of the best mode of the present invention, but the technical scope of the present invention is limited to the modes illustrated in the specific examples. Is not to be done.

(First embodiment)
In the method of wiping and removing a peroxide derivative-type explosive according to the first embodiment of the present invention described below, as a peroxide derivative-type explosive to be wiped off, a structure represented by the following formula (I) A trimeric acetone peroxide (TATP: 3,3,6,6,9,9-hexamethyl-1,2,4,5,7,6-hexaoxacyclononane) is selected.

(I) Synthesis of a dicarboxylic acid compound having the structure shown in formula (II) It has a ring structure having a characteristic ring structure and similarity to the trimeric acetone peroxide (TATP) of formula (I). A dicarboxylic acid compound having the structure represented by formula (II): 3- [12- (2-carboxyethyl) -9,12-dimethyl-7,8,10,11,13,14-hexaoxa-spiro- [5.8] Tetradec-9-yl] -propionic acid (3- [12- (2-carboxyethyl) -9,12-dimethyl-7,8,10,11,13,14-hexaoxa-spiro- [5.8] tetradec -9-yl] -propanoic acid). Hereinafter, the dicarboxylic acid compound having the structure represented by the formula (II) is abbreviated as TATP3.

  A method for synthesizing a dicarboxylic acid compound (TATP3) having a structure represented by the formula (II) has already been reported in the literature (Non-patent Document 2: Organic & Biomolecular Chemistry Vol. 4, p.4431-4436 (2006)). . A method for purifying a dicarboxylic acid compound (TATP3) having a structure represented by the formula (II) to be synthesized and a method for evaluating the purity thereof are also disclosed in this document.

  In the dicarboxylic acid compound (TATP3) having the structure represented by the formula (II), the spiro ring structure: carbon at the 9-position on the ring of 7,8,10,11,13,14-hexaoxa-spiro- [5.8] tetradecane Both the atom and the carbon atom at the 12-position are asymmetric centers (chiral centers). Regarding the configuration of the 9th carbon atom and the 12th carbon atom, the meso-type dicarboxylic acid compound (TATP3) that can be expressed as (9R, 12S) has a three-dimensional structure having two-fold rotational symmetry. Yes. Regarding the configuration of the carbon atom at the 9th position and the carbon atom at the 12th position, the cis-form dicarboxylic acid compound (TATP3) that can be expressed as (9R, 12R) has a three-dimensional structure having a symmetry plane.

  In the first embodiment, a mixture of the two stereoisomers is used. In addition, the dicarboxylic acid compound (TATP3) itself having a structure represented by the formula (II) does not exhibit immunogenicity.

(Ii) Immobilization of dicarboxylic acid compound (TATP3) having the structure shown in formula (II) on carrier protein Keyhole Limpet Hemocyanin is selected as the carrier protein.

  A modified carrier protein was prepared by binding a dicarboxylic acid compound (TATP3) represented by the formula (II) on the carrier protein by a carbodiimide method. In the first embodiment, the following two modified carrier proteins were prepared and used as immunogens for immunization.

(ii-a) Preparation of Modified Carrier Protein (TATP3-KLH-DMSO Immunogen) Using DMSO ((CH ₃ ) ₂ SO: Wako Pure Chemical Industries, Ltd.) as a reaction solvent, keyhole limpet hemocyanin ( A dicarboxylic acid compound (TATP3) represented by the formula (II) is bound on Keyhole Limpet Hemocyanin by the carbodiimide method to prepare a modified carrier protein (TATP3-KLH-DMSO immunogen). In the bond formation by the carbodiimide method, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC · HCl) (manufactured by Wako Pure Chemical Industries, Ltd.) is used as the binder carbodiimide.

  TATP3 dissolved in DMSO, 10 mg / 0.3 ml, and keyhole limpet hemocyanin (manufactured by Sigma Aldrich Japan) dissolved in DMSO, 10 mg / 1.7 ml are mixed. After mixing, the binder carbodiimide, 50 mg / 0.5 ml, dissolved in DMSO is added.

  A reaction solution using DMSO as a reaction solvent was allowed to stand at room temperature for 2 hours, and then allowed to stand at 4 ° C. for 12 hours to carry out the reaction. 0.1 ml of 1M glycine buffer (manufactured by Wako Pure Chemical Industries, Ltd.) adjusted to pH 8 was added to stop the reaction. The modified carrier protein (TATP3-KLH-DMSO immunogen) and the unreacted carrier protein contained in the solution were purified by dialysis with PBS (manufactured by Wako Pure Chemical Industries, Ltd.). A protein solution containing the prepared modified carrier protein (TATP3-KLH-DMSO immunogen) and unreacted carrier protein was used as a TATP3-DMSO (TATP3-KLH-DMSO immunogen) solution.

Under the reaction conditions using the above-described binder carbodiimide, the carboxyl group (—COOH) of the dicarboxylic acid compound (TATP3) represented by the formula (II) with respect to the amino group (—NH ₂ ) exposed on the surface of the carrier protein. Is used to form an amide bond (—CO—NH—), thereby binding the dicarboxylic acid compound (TATP3) represented by the formula (II).

  In addition, on the modified carrier protein (TATP3-KLH-DMSO immunogen) prepared under the above reaction conditions, the dicarboxylic acid compound (TATP3) represented by the formula (II) binds on average at 150 to 200 sites. is doing. That is, this corresponds to a state in which TATP3 is bound to the amino group of the lysine residue side chain on the surface of the carrier protein KLH.

(ii-b) Preparation of modified carrier protein (TATP3-KLH-borate buffer immunogen) Using a borate buffer at pH 8.5 as a reaction solvent, a formula on Keyhole Limpet Hemocyanin A dicarboxylic acid compound (TATP3) shown in (II) is bound by a carbodiimide method to prepare a modified carrier protein (TATP3-KLH-borate buffer immunogen). In the bond formation by the carbodiimide method, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC · HCl) (manufactured by Wako Pure Chemical Industries, Ltd.) is used as the binder carbodiimide. The pH 8.5 borate buffer is 0.992 g boric acid (Wako Pure Chemical Industries), 1.906 g borax (Wako Pure Chemical Industries), and 2.628 g NaCl (Wako Pure Chemical Industries). Is a buffer solution in which pH is adjusted to 8.5 by dissolving NaOH in 180 ml of pure water and adding NaOH.

  TATP3, 10 mg / (0.3 ml DMOS + 0.2 ml borate buffer) dissolved in a mixture of borate buffer and DMSO and keyhole limpet hemocyanin, 10 mg / 1.5 ml dissolved in borate buffer are mixed. After mixing, the binder carbodiimide, 50 mg / 0.5 ml, dissolved in borate buffer is added.

  A reaction solution using the borate buffer as a reaction solvent was allowed to stand at room temperature for 2 hours, and then allowed to stand at 4 ° C. for 12 hours to carry out the reaction. 0.1 ml of 1M glycine buffer (manufactured by Wako Pure Chemical Industries, Ltd.) adjusted to pH 8 was added to stop the reaction. The modified carrier protein (TATP3-KLH-DMSO immunogen) and the unreacted carrier protein contained in the solution were purified by dialysis with PBS (manufactured by Wako Pure Chemical Industries, Ltd.). A protein solution containing the prepared modified carrier protein (TATP3-KLH-borate buffer immunogen) and an unreacted carrier protein is converted into a TATP3-borate buffer (TATP3-KLH-borate buffer immunogen) solution. It was.

Under the reaction conditions using the above-described binder carbodiimide, the carboxyl group (—COOH) of the dicarboxylic acid compound (TATP3) represented by the formula (II) with respect to the amino group (—NH ₂ ) exposed on the surface of the carrier protein. Is used to form an amide bond (—CO—NH—), thereby binding the dicarboxylic acid compound (TATP3) represented by the formula (II).

  On the modified carrier protein (TATP3-KLH-borate buffer immunogen) prepared under the above reaction conditions, the dicarboxylic acid compound (TATP3) represented by the formula (II) averages 150 to 200. The place is connected.

(Iii) Preparation of Modified Carrier Protein for Antigen Modified with Dicarboxylic Acid Compound (TATP3) Represented by Formula (II) Bovine serum albumin (BSA) is selected as the carrier protein.

  A modified carrier protein was prepared by binding a dicarboxylic acid compound (TATP3) represented by the formula (II) on the carrier protein by a carbodiimide method. In the first embodiment, the following modified carrier protein was prepared and used as an antigen to be used for confirmation of antibody reactivity.

Preparation of Modified Carrier Protein for Antigen (TATP3-BSA Antigen) Using a borate buffer at pH 8.5 as a reaction solvent, a dicarboxylic acid compound (TATP3) represented by the formula (II) on bovine serum albumin (BSA) The modified carrier protein (TATP3-BSA antigen) is prepared by binding by the carbodiimide method. In the bond formation by the carbodiimide method, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC · HCl) (manufactured by Wako Pure Chemical Industries, Ltd.) is used as the binder carbodiimide. The pH 8.5 borate buffer is 0.992 g boric acid (Wako Pure Chemical Industries), 1.906 g borax (Wako Pure Chemical Industries), and 2.628 g NaCl (Wako Pure Chemical Industries). Is a buffer solution in which pH is adjusted to 8.5 by dissolving NaOH in 180 ml of pure water and adding NaOH.

  TATP3 dissolved in DMSO, 10 mg / 0.1 ml, bovine serum albumin (BSA) dissolved in pure water, 30 mg / 1.5 ml, and borate buffer 0.9 ml are mixed. After mixing, the binder carbodiimide, 50 mg / 0.25 ml, dissolved in borate buffer is added.

  A reaction solution using the borate buffer as a reaction solvent was allowed to stand at room temperature for 5 hours for reaction. 0.3 ml of 1M glycine buffer (manufactured by Wako Pure Chemical Industries, Ltd.) adjusted to pH 8 was added to stop the reaction. The modified carrier protein (TATP3-BSA antigen) and the unreacted carrier protein contained in the solution were purified by dialysis with PBS (manufactured by Wako Pure Chemical Industries, Ltd.). A protein solution containing the prepared modified carrier protein (TATP3-BSA antigen) and unreacted carrier protein was used as a TATP3-BSA (TATP3-BSA antigen) solution.

(Iv) Immunization using a modified carrier protein for an immunogen Sensitization is performed on the target non-human mammal using the above-mentioned two modified carrier proteins for an immunogen. In the first embodiment, a mouse (SLC: C57BL / 6) is selected as a mammal other than a human to be immunized.

  For immunization, a solution prepared by mixing the above-mentioned two modified immunogen carriers and proteins and adding Freund's complete adjuvant (Funakoshi Co., Ltd.) is used. The composition of the solution was 0.01 ml TATP3-DMSO (TATP3-KLH-DMSO immunogen) solution, 0.01 ml TATP3-borate buffer (TATP3-KLH-borate buffer immunogen) solution, 0.07 ml PBS and 0.01 ml of Freund's complete adjuvant (Funakoshi) are mixed uniformly.

  Sensitization (immune manipulation) was performed by subcutaneously injecting 1.0 ml of the solution into a 12-week-old mouse (SLC: C57BL / 6). On the 22nd, 35th, and 49th days after the first sensitization (first day), 1.0 ml of the solution was subcutaneously injected, and booster immunization was performed.

  On the 66th day after the first sensitization (first day), blood was collected from the immunized mice. Antiserum was prepared from the collected blood.

(V) Verification of cross-reactivity of polyclonal antibody contained in the obtained antiserum The polyclonal antibody contained in the obtained antiserum is a trimeric acetone peroxide (TATP) represented by the formula (I) It is verified using a competitive ELISA method that it shows cross-reactivity to.

  The polyclonal antibody contained in the obtained antiserum is presumed to be a mixture of two or more types of antibodies specific to the above-mentioned two modified immunogen carriers and proteins used for immunization. First, it is verified that an antibody exhibiting a specific reactivity with the dicarboxylic acid compound (TATP3) itself represented by the formula (II) is present in the polyclonal antibody.

  Specifically, as a carrier protein, the modified carrier protein for antigen (TATP3-BSA antigen) prepared using bovine serum albumin (BSA) instead of Keyhole Limpet Hemocyanin The presence of a reactive antibody is verified using an ELISA method.

  Using a TATP3-BSA (TATP3-BSA antigen) solution diluted 1000-fold, the TATP3-BSA (TATP3-BSA antigen) is immobilized on a plate for ELISA measurement by a natural adsorption method. After washing away and removing unadsorbed protein, 50 μl of PBS is added to the plate for ELISA measurement. Next, 50 μl of a solution obtained by diluting the obtained antiserum 100 times with PBS is added. The antibody is reacted with the TATP3-BSA (TATP3-BSA antigen) after standing at room temperature for 2 hours.

  After completion of the reaction, the solution on the plate for ELISA measurement is removed, and the plate is washed 3 times with 100 μl of each PBS. After the washing, 50 μl of an anti-mouse IgG-POD labeled antibody solution (Funakoshi) diluted 2000 times is added to the plate. The antibody that has been allowed to stand at room temperature for 1 hour and reacted with the antigen-modified carrier protein (TATP3-BSA antigen) on the plate is reacted with an anti-mouse IgG-POD-labeled antibody.

  After completion of the reaction, the solution on the plate for ELISA measurement is removed, and the plate is washed 4 times with 100 μl of each PBS. After the washing, 50 μl of ELISA peroxidase substrate (TMBZ, Funakoshi) solution is added to the plate. After carrying out an enzyme reaction for 60 minutes with the labeling enzyme peroxidase of the anti-mouse IgG-POD labeled antibody, 50 μl of 1N sulfuric acid (manufactured by Wako Pure Chemical Industries, Ltd.) is added to stop the reaction. The concentration of the reaction product by the enzyme reaction is determined by measuring the absorbance at 450 nm. In FIG. 1, the measurement result denoted as “buffer” corresponds to the amount of antibody reacted with TATP3-BSA (TATP3-BSA antigen).

  The above-mentioned antibody having reactivity with TATP3-BSA (TATP3-BSA antigen) reacts with the dicarboxylic acid compound (TATP3) itself represented by the formula (II) bound on bovine serum albumin (BSA). It is an antibody. Therefore, it was verified that the polyclonal antibody contained in the obtained antiserum contains an antibody having a specific reactivity with the dicarboxylic acid compound (TATP3) itself represented by the formula (II).

  Next, among the plurality of types of antibodies reactive to the dicarboxylic acid compound (TATP3) itself represented by the formula (II) contained in the obtained antiserum polyclonal antibody, the trimeric compound represented by the formula (I) The presence of an antibody showing cross-reactivity to the body form of acetone peroxide (TATP) is verified using a competitive ELISA method.

  Using a TATP3-BSA (TATP3-BSA antigen) solution diluted 1000-fold, the TATP3-BSA (TATP3-BSA antigen) is immobilized on a plate for ELISA measurement by a natural adsorption method. After washing and removing unadsorbed protein, 50 μl of TATP (AQ Standard) PBS solution is added to the ELISA measurement plate so that the final concentration is 100 ppm. Next, 50 μl of a solution obtained by diluting the obtained antiserum 100 times with PBS is added. The antibody is reacted with the TATP3-BSA (TATP3-BSA antigen) after standing at room temperature for 2 hours. The final concentration of 100 ppm of TATP (molecular weight 222.4) contained in the reaction solution corresponds to 0.45 mM.

  When the antigen-antibody reaction proceeds between TATP and the antibody added to the solution on the plate during the reaction, TATP3-BSA immobilized on the plate (TATP3-BSA antigen) and Competition with the antigen-antibody reaction. As a result, the amount of antibody immobilized on the plate is reduced through an antigen-antibody reaction with TATP3-BSA (TATP3-BSA antigen) immobilized on the plate.

  After completion of the reaction, the solution on the plate for ELISA measurement is removed, and the plate is washed 4 times with 100 μl of each PBS. After the washing, 50 μl of ELISA peroxidase substrate (TMBZ, Funakoshi) solution is added to the plate. After carrying out an enzyme reaction for 60 minutes with the labeling enzyme peroxidase of the anti-mouse IgG-POD labeled antibody, 50 μl of 1N sulfuric acid (manufactured by Wako Pure Chemical Industries, Ltd.) is added to stop the reaction. The concentration of the reaction product by the enzyme reaction is determined by measuring the absorbance at 450 nm.

  Moreover, it replaces with the PBS solution of TATP, PBS is added, and it reacts similarly.

  Using a TATP3-BSA (TATP3-BSA antigen) solution diluted 1000-fold, the TATP3-BSA (TATP3-BSA antigen) is immobilized on a plate for ELISA measurement by a natural adsorption method. After washing away and removing unadsorbed protein, 50 μl of PBS is added to the plate for ELISA measurement. Next, 50 μl of a solution obtained by diluting the obtained antiserum 100 times with PBS is added. The antibody is reacted with the TATP3-BSA (TATP3-BSA antigen) after standing at room temperature for 2 hours.

  After completion of the reaction, the solution on the plate for ELISA measurement is removed, and the plate is washed 4 times with 100 μl of each PBS. After the washing, 50 μl of ELISA peroxidase substrate (TMBZ, Funakoshi) solution is added to the plate. After carrying out an enzyme reaction for 60 minutes with the labeling enzyme peroxidase of the anti-mouse IgG-POD labeled antibody, 50 μl of 1N sulfuric acid (manufactured by Wako Pure Chemical Industries, Ltd.) is added to stop the reaction. The concentration of the reaction product by the enzyme reaction is determined by measuring the absorbance at 450 nm.

  The absorbance at 450 nm measured when the TATP solution in PBS is added is compared with the absorbance measured at 450 nm when PBS is added. As a result, it was confirmed that when the PBS solution of TATP was added, the absorbance at 450 nm was reduced as compared with the case where PBS was added. That is, it was confirmed that antibodies having cross-reactivity with TATP exist in the obtained antiserum.

  Therefore, it was verified that the obtained antiserum polyclonal antibody contains an antibody exhibiting cross-reactivity with the trimeric acetone peroxide (TATP) represented by the formula (I).

  The anti-mouse IgG-POD-labeled antibody has specificity for the mouse IgG1-type antibody, and is an antibody that exhibits cross-reactivity with the trimeric acetone peroxide (TATP) represented by the formula (I). The type was confirmed to be IgG1 type.

(Vi) Creation of a mono-normal antibody against a dicarboxylic acid compound (TATP3) having the structure shown in formula (II) In the obtained antiserum polyclonal antibody, trimeric acetone peroxide (TATP) shown in formula (I) is contained. Cross-reactivity to trimeric acetone peroxide (TATP) represented by formula (I) using a group of mouse antibody-producing cells that have been verified to contain antibodies that exhibit cross-reactivity to Hybridoma cells producing a monoclonal antibody exhibiting

  After the first sensitization (first day) after this verification, the spleen is extracted from the mouse on the 66th day, and spleen cells are prepared.

The prepared mouse spleen cells and P3-X63-Ag8-U mouse myeloma cells in a ratio of 5: 1 cells in RPMI 1640 medium (manufactured by Invitrogen), 50% polyethylene glycol (sum) In the presence of Kogyo Pharmaceutical Co., Ltd.), the mixture is mixed at 37 ° C. for 2 minutes to cause cell fusion. After the cell fusion treatment, the resulting hybridoma cells are suspended in a HAT medium (20% fetal bovine serum) and then dispensed into a microplate. The hybridoma cells dispensed on the microplate are cultured in a carbon dioxide incubator at 37 ° C. and 5% CO ₂ . During the culture, half of the medium is replaced with fresh HT medium (10% fetal calf serum) once every four days.

  The HAT medium is obtained by adding an appropriate amount of HAT supplement (manufactured by Invitrogen) to RPMI 1640 medium. In the first embodiment, 20 μl of HAT supplement is added per 1 ml of RPMI 1640 medium.

  The HT medium is obtained by adding an appropriate amount of HT supplement (manufactured by Invitrogen) to RPMI1640 medium. In the first embodiment, 20 μl of HT supplement is added per 1 ml of RPMI 1640 medium.

  Under the above culture conditions, the hybridoma cells dispensed on the microplate were cultured for 2 weeks to establish hybridoma cell lines, respectively.

  Next, whether or not the monoclonal antibody produced in the culture supernatant of each hybridoma cell line is a monoclonal antibody having a specific reactivity with the dicarboxylic acid compound (TATP3) itself represented by the formula (II) Was confirmed. Furthermore, among those that have been verified to be a monoclonal antibody having specific reactivity with the dicarboxylic acid compound (TATP3) itself represented by the formula (II), a trimeric acetone peroxide represented by the formula (I) ( Antibodies showing cross-reactivity to TATP) were selected.

(vi-a) Verification of whether or not the dicarboxylic acid compound (TATP3) itself represented by the formula (II) is a monoclonal antibody having specific reactivity Specifically, in the culture supernatant of each hybridoma cell line Whether the monoclonal antibody contained in the antibody is a monoclonal antibody having reactivity with the modified carrier protein for antigen (TATP3-BSA antigen) prepared using bovine serum albumin (BSA) is determined by ELISA. Use and verify.

  Using a TATP3-BSA (TATP3-BSA antigen) solution diluted 1000-fold, the TATP3-BSA (TATP3-BSA antigen) is immobilized on a plate for ELISA measurement by a natural adsorption method. After washing away and removing unadsorbed protein, 50 μl of PBS is added to the plate for ELISA measurement. Next, 50 μl of a solution obtained by diluting the culture supernatant of each hybridoma cell line 100 times with PBS is added. The mixture is allowed to stand at room temperature for 2 hours, and the monoclonal antibody is reacted with the TATP3-BSA (TATP3-BSA antigen).

  After completion of the reaction, the solution on the plate for ELISA measurement is removed, and the plate is washed 3 times with 100 μl of each PBS. After the washing, 50 μl of an anti-mouse IgG-POD labeled antibody solution (Funakoshi) diluted 2000 times is added to the plate. The antibody that has been allowed to stand at room temperature for 1 hour and reacted with the antigen-modified carrier protein (TATP3-BSA antigen) on the plate is reacted with an anti-mouse IgG-POD-labeled antibody.

  After completion of the reaction, the solution on the plate for ELISA measurement is removed, and the plate is washed 4 times with 100 μl of each PBS. After the washing, 50 μl of ELISA peroxidase substrate (TMBZ, Funakoshi) solution is added to the plate. After carrying out an enzyme reaction for 60 minutes with the labeling enzyme peroxidase of the anti-mouse IgG-POD labeled antibody, 50 μl of 1N sulfuric acid (manufactured by Wako Pure Chemical Industries, Ltd.) is added to stop the reaction. The concentration of the reaction product by the enzyme reaction is determined by measuring the absorbance at 450 nm.

  The monoclonal antibody having reactivity to the above TATP3-BSA (TATP3-BSA antigen) reacts with the dicarboxylic acid compound (TATP3) itself represented by the formula (II) bound on bovine serum albumin (BSA). It is a monoclonal antibody.

  According to this screening procedure, it was verified whether or not a monoclonal antibody having specific reactivity with the dicarboxylic acid compound (TATP3) itself represented by the formula (II) exists in the culture supernatant of each hybridoma cell line. . As a result, it was confirmed that, out of a total of 13 clones of the hybridoma cell line, 13 clones produced monoclonal antibodies that reacted with the dicarboxylic acid compound (TATP3) itself represented by the formula (II).

(vi-b) Selection of monoclonal antibody showing cross-reactivity to trimeric acetone peroxide (TATP) represented by formula (I) Formula (II) selected by primary screening (vi-a) Using a competitive ELISA method, a monoclonal antibody exhibiting cross-reactivity to trimeric acetone peroxide (TATP) represented by formula (I) is selected from a plurality of monoclonal antibodies reactive to the dicarboxylic acid compound (TATP3) itself shown. And sort.

  Using a TATP3-BSA (TATP3-BSA antigen) solution diluted 1000-fold, the TATP3-BSA (TATP3-BSA antigen) is immobilized on a plate for ELISA measurement by a natural adsorption method. After washing away and removing unadsorbed protein, 50 μl of TATP in PBS is added to the ELISA measurement plate to a final concentration of 100 ppm. Subsequently, 50 μl of a solution obtained by diluting the culture supernatant of each selected hybridoma cell line 100 times with PBS is added. The antibody is reacted with the TATP3-BSA (TATP3-BSA antigen) after standing at room temperature for 2 hours. A final concentration of 100 ppm of TATP contained in the reaction solution corresponds to 0.45 mM.

  When the antigen-antibody reaction proceeds between TATP and the antibody added to the solution on the plate during the reaction, TATP3-BSA immobilized on the plate (TATP3-BSA antigen) and Competition with the antigen-antibody reaction. As a result, the amount of antibody immobilized on the plate is reduced through an antigen-antibody reaction with TATP3-BSA (TATP3-BSA antigen) immobilized on the plate.

  After completion of the reaction, the solution on the plate for ELISA measurement is removed, and the plate is washed 4 times with 100 μl of each PBS. After the washing, 50 μl of ELISA peroxidase substrate (TMBZ, Funakoshi) solution is added to the plate. After carrying out an enzyme reaction for 60 minutes with the labeling enzyme peroxidase of the anti-mouse IgG-POD labeled antibody, 50 μl of 1N sulfuric acid (manufactured by Wako Pure Chemical Industries, Ltd.) is added to stop the reaction. The concentration of the reaction product by the enzyme reaction is determined by measuring the absorbance at 450 nm.

  Compared to the case where TATP is not added to the solution on the plate during the antigen-antibody reaction, when TATP is added, a result indicating that the concentration of the reaction product due to the enzyme reaction decreases is obtained. When added, it is judged that competition has occurred. That is, when such competition occurs, the monoclonal antibody contained in the culture supernatant of the hybridoma cell line can be determined as a monoclonal antibody exhibiting cross-reactivity with TATP.

  In accordance with this screening procedure, from a plurality of monoclonal antibodies reactive to the dicarboxylic acid compound (TATP3) itself represented by the formula (II), selected in the primary screening of (vi-a), the trimer represented by the formula (I) Monoclonal antibodies showing cross-reactivity with body type acetone peroxide (TATP) were selected.

  As a result, a plurality of types of monoclonal antibodies showing cross-reactivity with the trimeric acetone peroxide (TATP) represented by the formula (I) were selected. FIG. 1 shows, as an example, measurement results of four monoclonal antibodies: mAb-T001 to mAb-T004 among the plurality of monoclonal antibodies selected by this secondary screening.

  In FIG. 1, the measurement result expressed as “Borate buffer” is the measurement result by the ELISA method in the primary screening (vi-a), that is, TATP3-BSA (TATP3-BSA) in the situation where TATP does not exist. This corresponds to the amount of the monoclonal antibody reacted against the antigen). In FIG. 1, the measurement result expressed as “100 ppm TATP” corresponds to the amount of monoclonal antibody reacted with TATP3-BSA (TATP3-BSA antigen) in the situation where the above-mentioned TATP coexists. Yes.

  The results shown in FIG. 1 show that TATP3 immobilized on the plate because the antigen-antibody reaction proceeds between TATP and the monoclonal antibody added to the solution on the plate during the above reaction. -It clearly shows that an antigen-antibody reaction with BSA (TATP3-BSA antigen) and competition have occurred. That is, the four types of monoclonal antibodies are monoclonal antibodies that are reactive with the dicarboxylic acid compound (TATP3) itself represented by the formula (II), and further, trimeric acetone peroxide (TATP) represented by the formula (I). It is the result of verifying that it is also a monoclonal antibody showing cross-reactivity to).

  Therefore, the monoclonal antibody selected by the primary screening of (vi-a) and the secondary screening of (vi-b) is cross-linked to trimeric acetone peroxide (TATP) represented by formula (I). It was confirmed to be a monoclonal antibody exhibiting reactivity.

In addition, the result shown in FIG. 1 shows that when a trimeric acetone peroxide (TATP) represented by the formula (I) is added to a reaction solution for performing an antigen-antibody reaction to a final concentration of 0.45 mM,
In mAb-T001 to mAb-T004, the result suggests that at least about 10% is bound to TATP.

  Subsequently, a method for immobilizing the monoclonal antibody on the water-absorbing material will be described below.

  In the first embodiment, when the monoclonal antibody is immobilized, a water-absorbing material composed of a highly liquid-absorbing / solvent-resistant cellulose nonwoven fabric is used as a liquid-absorbing material having a liquid-absorbing property.

  Specifically, BEMCOT high-grade wiper M-3II manufactured by Asahi Kasei Fibers Co., Ltd., which is an example of a highly liquid-absorbing and solvent-resistant cellulose nonwoven fabric, is employed as the water-absorbing material. The cellulose non-woven fabric is immersed in 16 ml of a monoclonal antibody: mAb-T003 solution having a concentration of 15 mg / ml for 1 hour. During the immersion treatment, the monoclonal antibody is non-selectively adsorbed on the surface of the fibers constituting the cellulose nonwoven fabric. The density of the adsorbed monoclonal antibody is proportional to the concentration of the monoclonal antibody in the solution.

  The amount of the monoclonal antibody adsorbed non-selectively is in the range of 220 mg to 240 mg per 5 cm × 5 cm of the cellulose nonwoven fabric.

  After pulling up from the monoclonal antibody solution, the solution is left at room temperature for 1 hour to evaporate water. In the process of evaporating moisture, the evaporation of moisture proceeds from the upper surface side of the cellulose nonwoven fabric, while the lower surface side is further infiltrated. As the moisture evaporates, the concentration of the remaining monoclonal antibody solution proceeds, and the density of the monoclonal antibody adsorbed on the lower surface side of the cellulose nonwoven fabric is higher than that on the upper surface side.

  Then, it is immersed for 1 hour in an aqueous solution of polyvinyl alcohol having a concentration of 1 w / v%. The surface of the fiber which comprises this cellulose nonwoven fabric will be in the state coat | covered with polyvinyl alcohol. At the same time, the monoclonal antibody molecules adsorbed on the fiber surface are also covered with polyvinyl alcohol.

  In addition to the monoclonal antibody molecules adsorbed on the surface of the fiber, the monoclonal antibody dissolved in the solution of the monoclonal antibody remained in the cellulose nonwoven fabric. When immersed in an aqueous solution of polyvinyl alcohol, the remaining monoclonal antibody is redissolved in the aqueous solution. The re-dissolved monoclonal antibody molecule is also partially covered with polyvinyl alcohol. Therefore, the frequency of elution of the monoclonal antibody molecules coated with polyvinyl alcohol from the inside of the cellulose nonwoven fabric by diffusion is remarkably reduced.

  Therefore, the inside of the cellulose nonwoven fabric is maintained in a state substantially filled with the monoclonal antibody solution. In this state, desorption of the monoclonal antibody molecules adsorbed on the fiber surface does not proceed, and an adsorbed layer of polyvinyl alcohol is formed so as to cover the adsorbed monoclonal antibody molecules and the fiber surface.

  After pulling up from the aqueous solution of polyvinyl alcohol, it is allowed to stand at room temperature to evaporate the contained water and dry sufficiently. When the drying process is performed, the removal of solvated water molecules contained in the adsorption layer of polyvinyl alcohol formed so as to cover the surface of the adsorbed monoclonal antibody molecules and fibers proceeds, and the polyvinyl alcohol coating layer is formed. Converted.

  The wiping treatment material finally produced is polyvinyl alcohol so that the monoclonal antibody molecules are adsorbed on the surface of the fibers constituting the cellulose nonwoven fabric of the base material, and the surface of the adsorbed monoclonal antibody molecules and the fibers are covered. The coating layer is formed.

  Using the wiping treatment material produced by the above procedure, it was verified that the solid powder of trimeric acetone peroxide can be wiped off.

  Trimeric acetone peroxide microcrystal powder is prepared by using 0.01% solution of AccuStandard as a trimeric acetone peroxide solution.

  5 ml of the 0.01% solution is dropped on a glass substrate and evaporated to dryness at room temperature. The contained 0.5 mg of trimeric acetone peroxide is precipitated as fine crystals. A number of glass substrates on which the trimeric acetone peroxide microcrystal powder was deposited were prepared, and a comparative experiment was performed under the following three conditions.

(Condition 1)
In the wiping treatment material, acetonitrile / water mixed solution dissolved in pure water at a concentration of 5 v / v%, 10 v / v%, 25 v / v%, 50 v / v%, and acetonitrile (100 v / v%), After absorbing 2 ml of each to make it infiltrated, the trimeric acetone peroxide microcrystal powder on the glass substrate is covered. In this state, a total of 0.5 mg of trimeric acetone peroxide microcrystal powder is dissolved with an infiltrating acetonitrile / water mixture or acetonitrile, and then the wiping treatment material is removed.

  After finishing the wiping process, the surface of the glass substrate is observed.

  When the acetonitrile concentration is 5 v / v%, 10 v / v%, 25 v / v%, and 50 v / v% in the range observed visually, the trimer type is formed on the glass substrate. The residue of fine crystals of acetone peroxide was not confirmed. That is, after the wiping treatment, a slight film layer of the solution remains on the glass substrate, but when the solvent in the remaining film layer of the solution evaporates, a fine crystalline residue of trimeric acetone peroxide is left. It was not found.

  On the other hand, when acetonitrile (100 v / v%) is used, after the wiping treatment, a slight film layer of the solution remains on the glass substrate, and when the solvent in the remaining film layer of the solution evaporates, A fine crystalline residue of monomeric acetone peroxide was found.

(Condition 2)
A 1: 1 (v / v) mixed solvent of methanol and acetonitrile was soaked in pure water at 5 v / v%, 10 v / v so that 0.5 mg of trimeric acetone peroxide microcrystalline powder on a glass substrate was immersed. 2 ml each of (methanol / acetonitrile) / water mixed solution mixed at a ratio of 25%, 25v / v%, 50v / v%, or 1: 1 (v / v) mixed solvent of methanol and acetonitrile is dropped. When the microcrystalline powder is dissolved by the mixed solution and the mixed solvent, the solution is covered with the wiping treatment material. It is confirmed that the solution penetrates into the covered wiping treatment material, is sucked into the wiping treatment material, and becomes infiltrated. At that point, the wiping treatment material is removed.

  After finishing the wiping process, the surface of the glass substrate is observed.

  In the range observed visually, when a (methanol / acetonitrile) / water mixture having a mixing ratio of 5 v / v%, 10 v / v%, and 25 v / v% is used, a trimer type is formed on the glass substrate. The residue of fine crystals of acetone peroxide was not confirmed. That is, after the wiping treatment, a slight film layer of the solution remains on the glass substrate, but when the solvent in the remaining film layer of the solution evaporates, a fine crystalline residue of trimeric acetone peroxide is left. It was not found.

  On the other hand, even when a 1: 1 (v / v) mixed solvent of methanol and acetonitrile and a (methanol / acetonitrile) / water mixed solution with a mixing ratio of 50 v / v% are used, after wiping treatment, the glass substrate A slight film layer of solution remains on top. When the solvent in the film layer of the remaining solution was evaporated, a residue of fine crystals of trimeric acetone peroxide was found.

(Condition 3)
To the wiping treatment material, an acetone / water mixture solution dissolved in pure water at a concentration of 5 v / v%, 10 v / v%, 25 v / v%, 50 v / v%, and acetone (100 v / v%), After absorbing 2 ml of each to make it infiltrated, the trimeric acetone peroxide microcrystal powder on the glass substrate is covered. In this state, a total of 0.5 mg of trimeric acetone peroxide microcrystal powder is dissolved with an infiltrated acetone / water mixture or acetone, and then the wiping treatment material is removed.

  After finishing the wiping process, the surface of the glass substrate is observed.

  In the range observed visually, when an acetone / water mixture having an acetone concentration of 5 v / v%, 10 v / v%, and 25 v / v% is used, fine trimer-type acetone peroxide is formed on the glass substrate. No residual crystal was confirmed. That is, after the wiping treatment, a slight film layer of the solution remains on the glass substrate, but when the solvent in the remaining film layer of the solution evaporates, a fine crystalline residue of trimeric acetone peroxide is left. It was not found.

  On the other hand, even when acetone (100 v / v%) or an acetone / water mixed solution having an acetone concentration of 50 v / v% is used, a film layer of the solution slightly remains on the glass substrate after the wiping treatment. . When the solvent in the film layer of the remaining solution was evaporated, a residue of fine crystals of trimeric acetone peroxide was found.

(Condition 4)
To the above-mentioned wiping treatment material, an ethanol / water mixture solution dissolved in pure water at a concentration of 5 v / v%, 10 v / v%, 25 v / v%, 50 v / v%, and ethanol (100 v / v%), After absorbing 2 ml of each to make it infiltrated, the trimeric acetone peroxide microcrystal powder on the glass substrate is covered. In that state, a total of 0.5 mg of trimeric acetone peroxide microcrystal powder is dissolved with an infiltrated ethanol / water mixture or ethanol, and then the wiping treatment material is removed.

  After finishing the wiping process, the surface of the glass substrate is observed.

  In the range observed visually, no residue of fine crystals of trimeric acetone peroxide was observed on the glass substrate when either ethanol / water mixture or ethanol was used. That is, after the wiping treatment, a slight film layer of the solution remains on the glass substrate, but when the solvent in the remaining film layer of the solution evaporates, a fine crystalline residue of trimeric acetone peroxide is left. It was not found.

(Condition 5)
An ethanolamine / water mixture solution dissolved in pure water at concentrations of 10 v / v%, 25 v / v%, 50 v / v%, 80 v / v%, and ethanolamine (100 v / v%) 2 ml of each is absorbed to make it infiltrated, and the trimeric acetone peroxide microcrystal powder on the glass substrate is covered. In this state, a total of 0.5 mg of trimeric acetone peroxide microcrystal powder is dissolved with an infiltrated ethanolamine / water mixture or ethanolamine, and then the wiping treatment material is removed.

  After finishing the wiping process, the surface of the glass substrate is observed.

  When the ethanolamine concentration is 10 v / v%, 25 v / v%, 50 v / v% in the range observed visually, an ethanolamine / water mixture is used. The residue of fine crystals of acetone was not confirmed. That is, after the wiping treatment, a slight film layer of the solution remains on the glass substrate, but when the solvent in the remaining film layer of the solution evaporates, a fine crystalline residue of trimeric acetone peroxide is left. It was not found.

  On the other hand, even when ethanolamine (100 v / v%) or an ethanolamine / water mixture having an ethanolamine concentration of 80 v / v% is used, a slightly film of the solution is formed on the glass substrate after the wiping treatment. The layer remains. When the solvent in the film layer of the remaining solution was evaporated, a residue of fine crystals of trimeric acetone peroxide was found.

(Condition 6)
A methyl ethyl ketone / water mixture solution dissolved in pure water at a concentration of 5 v / v%, 10 v / v%, 25 v / v%, 50 v / v%, 80 v / v%, and methyl ethyl ketone (100 v / v 2% of each is absorbed so as to be infiltrated, and the trimeric acetone peroxide microcrystal powder on the glass substrate is covered. In this state, a total of 0.5 mg of trimeric acetone peroxide microcrystal powder is dissolved with the infiltrated methyl ethyl ketone / water mixture or methyl ethyl ketone, and then the wiping treatment material is removed.

  After finishing the wiping process, the surface of the glass substrate is observed.

  In the range observed visually, when a methyl ethyl ketone / water mixture having a methyl ethyl ketone concentration of 5 v / v%, 10 v / v%, and 25 v / v% is used, fine trimer type acetone peroxide is formed on the glass substrate. No residual crystal was confirmed. That is, after the wiping treatment, a slight film layer of the solution remains on the glass substrate, but when the solvent in the remaining film layer of the solution evaporates, a fine crystalline residue of trimeric acetone peroxide is left. It was not found.

  On the other hand, even when methyl ethyl ketone (100 v / v%) or a methyl ethyl ketone / water mixture having a methyl ethyl ketone concentration of 50 v / v% or 80 v / v% is used, a slight amount of A film layer of solution remains. When the solvent in the film layer of the remaining solution was evaporated, a residue of fine crystals of trimeric acetone peroxide was found.

(Condition 7)
Wiping is performed using the cellulose nonwoven fabric of the base material that has not been subjected to the above-described monoclonal antibody adsorption treatment.

  The cellulose non-woven fabric is absorbed with 2 ml of acetonitrile / water mixture dissolved in pure water at a concentration of 50 v / v% to make it infiltrated, and then trimeric acetone peroxide microcrystal powder on a glass substrate Cover. In this state, a total of 0.5 mg of trimeric acetone peroxide microcrystal powder is dissolved by an infiltrating acetonitrile / water mixture, and then the cellulose nonwoven fabric is removed.

  After finishing the wiping process, the surface of the glass substrate is observed.

  In the range observed visually, the residue of fine crystals of trimeric acetone peroxide was confirmed on the glass substrate. That is, when the cellulose nonwoven fabric is removed, a film layer of the solution slightly remains on the glass substrate, but the solvent in the film layer of the remaining solution evaporates, and trimeric acetone peroxide fine crystals are formed. A residue was found.

  In the above-mentioned conditions 1 to 7, when a wiping treatment material or the cellulose nonwoven fabric is removed, a phenomenon that a film layer of a liquid slightly remains on the glass substrate has been found similarly.

  Of course, in condition 7, the film layer of the remaining liquid is a solution in which trimeric acetone peroxide is dissolved at a concentration of 0.5 mg / 2 ml in an acetonitrile / water mixture.

Also,
In condition 1, when acetonitrile (100 v / v%) is used;
In condition 2, when a 1: 1 (v / v) mixed solvent of methanol and acetonitrile and a (methanol / acetonitrile) / water mixed solution with a mixing ratio of 50 v / v% are used;
In condition 3, when using acetone (100 v / v%) or an acetone / water mixture having an acetone concentration of 50 v / v%;
In condition 5, when using ethanolamine (100 v / v%) or an ethanolamine / water mixture having an ethanolamine concentration of 80 v / v%; and
In condition 5, even when methyl ethyl ketone (100 v / v%) or a methyl ethyl ketone / water mixture having a methyl ethyl ketone concentration of 50 v / v% and 80 v / v% is used, It is judged that each of the mixed solvents is a solution in which trimeric acetone peroxide is dissolved in a considerable concentration. The concentration is considerably reduced compared with the initial concentration of 0.5 mg / 2 ml, but when the solvent in the remaining film layer of the solution evaporates, the trimeric acetone peroxide has a fine concentration. The concentration is sufficient to produce a crystalline residue.

on the other hand,
In condition 1, when an acetonitrile / water mixture having an acetonitrile concentration of 5 v / v%, 10 v / v%, 25 v / v%, 50 v / v% is used;
In condition 2, when a (methanol / acetonitrile) / water mixture having a mixing ratio of 5 v / v%, 10 v / v%, and 25 v / v% is used;
In condition 3, when an acetone / water mixture having an acetone concentration of 5 v / v%, 10 v / v%, and 25 v / v% is used;
When the ethanol concentration is 5 v / v%, 10 v / v%, 25 v / v%, 50 v / v%, or ethanol (100 v / v%) in condition 4;
In condition 5, when an ethanolamine / water mixture having an ethanolamine concentration of 10 v / v%, 25 v / v%, 50 v / v% is used; and
In condition 6, when a methyl ethyl ketone / water mixed solution having a methyl ethyl ketone concentration of 5 v / v%, 10 v / v%, or 25 v / v% is used, the film layer of the slightly remaining liquid has a trimer type excess. The liquid is judged to be substantially free of acetone oxide.

  Therefore, the acetonitrile / water mixture, the (methanol / acetonitrile) / water mixture, the acetone / water mixture, the ethanol / water mixture, the ethanolamine / water mixture, the methyl ethyl ketone / water mixture with the above mixing ratio are used, It was verified that the trimeric acetone peroxide microcrystal powder on the glass substrate could be efficiently wiped off by using the wiping treatment material on which the monoclonal antibody was immobilized.

(Second embodiment)
Also in the second embodiment, a wiping treatment material is produced using a monoclonal antibody: mAb-T003.

  A method for immobilizing the monoclonal antibody to the water-absorbing material in the wiping treatment material produced in the second embodiment will be described below.

  In the second embodiment, when immobilizing a monoclonal antibody, as a liquid-absorbing material having a liquid-absorbing property as a base material, a hyaluronic acid excellent in water retention and carboxymethyl cellulose as components are used in a film form. A water-absorbing material, specifically, Sepurafilm (registered trademark) of Genzyme is used.

  A 5 cm x 5 cm film of Sepurafilm (R) from Genzyme is used. In advance, activation treatment of hyaluronic acid and carboxyl groups present in carboxymethylcellulose constituting the film-like water-absorbing material is performed. In the carboxyl group activation treatment, carbodiimide and N-hydroxysuccinimide are used. As the carbodiimide, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC · HCl) (manufactured by Wako Pure Chemical Industries, Ltd.) is used.

  Specifically, the film was immersed in 3 ml of an activation treatment solution containing 0.2 M 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride and 0.05 M N-hydroxysuccinimide. Let stand at room temperature for 1 hour. After the activation treatment, the film taken out from the activation treatment solution is immersed in physiological saline containing a phosphate buffer, and the activation treatment solution infiltrated in the film is sufficiently diluted and washed.

  Next, protein A is allowed to act on the activated carboxyl group to immobilize protein A.

  The film subjected to the carboxyl group activation treatment is immersed in 7 ml of a protein A gel (PIERCE, IL, USA) and allowed to stand at room temperature for 1 hour. After the immobilization treatment, the film taken out from the protein A gel solution is immersed in physiological saline containing a phosphate buffer and thoroughly washed. This protein A gel can immobilize 35 mg of IgG antibody per ml.

  Under the above-described immobilization conditions, the total amount of antibody that can be immobilized by protein A immobilized on a 5 cm × 5 cm film is about 240 mg.

  Next, the protein A-immobilized film is immersed in 16 ml of a solution of monoclonal antibody: mAb-T003 having a concentration of 15 mg / ml for 2 hours. Monoclonal antibody: mAb-T003 is bound to protein A, and is immobilized on the substrate Seprafilm (registered trademark) using the protein A as a linker.

  The total amount of monoclonal antibody immobilized on a 5 cm × 5 cm film is approximately 240 mg.

  Then, it is immersed for 1 hour in an aqueous solution of polyvinyl alcohol having a concentration of 1 w / v%. The outer surface and the entire inner surface of the film are covered with polyvinyl alcohol. At the same time, the monoclonal antibody molecules immobilized on the substrate Seprafilm (registered trademark) using the protein A as a linker are also coated with polyvinyl alcohol.

  After pulling up from the aqueous solution of polyvinyl alcohol, it is allowed to stand at room temperature to evaporate the contained water and dry sufficiently. When the drying treatment is performed, removal of solvated water molecules contained in the adsorption layer of polyvinyl alcohol formed so as to cover the surface of the immobilized monoclonal antibody molecule and the substrate film proceeds, and the polyvinyl alcohol It is converted into a coating layer.

  Finally, the wiping treatment material is made of a polyvinyl alcohol coating layer in which the monoclonal antibody molecules are immobilized on the film surface of the substrate, and the immobilized monoclonal antibody molecules and the substrate film surface are covered. Is formed.

  Using the wiping treatment material produced by the above procedure, it was verified that the solid powder of trimeric acetone peroxide can be wiped off.

  Trimeric acetone peroxide microcrystal powder is prepared by using 0.01% solution of AccuStandard as a trimeric acetone peroxide solution.

  5 ml of the 0.01% solution is dropped on a glass substrate and evaporated to dryness at room temperature. The contained 0.5 mg of trimeric acetone peroxide is precipitated as fine crystals. A number of glass substrates on which the trimeric acetone peroxide microcrystal powder was deposited were prepared, and a comparative experiment was performed under the following three conditions.

(Condition 1)
In the wiping treatment material, acetonitrile / water mixed solution dissolved in pure water at a concentration of 5 v / v%, 10 v / v%, 25 v / v%, 50 v / v%, and acetonitrile (100 v / v%), After absorbing 2 ml of each to make it infiltrated, the trimeric acetone peroxide microcrystal powder on the glass substrate is covered. In this state, a total of 0.5 mg of trimeric acetone peroxide microcrystal powder is dissolved with an infiltrating acetonitrile / water mixture or acetonitrile, and then the wiping treatment material is removed.

  After finishing the wiping process, the surface of the glass substrate is observed.

  When the acetonitrile concentration is 5 v / v%, 10 v / v%, 25 v / v%, and 50 v / v% in the range observed visually, the trimer type is formed on the glass substrate. The residue of fine crystals of acetone peroxide was not confirmed. That is, after the wiping treatment, a slight film layer of the solution remains on the glass substrate, but when the solvent in the remaining film layer of the solution evaporates, a fine crystalline residue of trimeric acetone peroxide is left. It was not found.

  On the other hand, when acetonitrile (100 v / v%) is used, a film layer of the solution slightly remains on the glass substrate after the wiping treatment, but when the solvent in the film layer of the remaining solution evaporates, A fine crystalline residue of trimeric acetone peroxide was found.

(Condition 2)
A 1: 1 (v / v) mixed solvent of methanol and acetonitrile was soaked in pure water at 5 v / v%, 10 v / v so that 0.5 mg of trimeric acetone peroxide microcrystalline powder on a glass substrate was immersed. 2 ml each of (methanol / acetonitrile) / water mixed solution mixed at a ratio of 25%, 25v / v%, 50v / v%, or 1: 1 (v / v) mixed solvent of methanol and acetonitrile is dropped. When the microcrystalline powder is dissolved by the mixed solution and the mixed solvent, the solution is covered with the wiping treatment material. It is confirmed that the solution penetrates into the covered wiping treatment material, is sucked into the wiping treatment material, and becomes infiltrated. At that point, the wiping treatment material is removed.

  After finishing the wiping process, the surface of the glass substrate is observed.

  In the range observed visually, when a (methanol / acetonitrile) / water mixture having a mixing ratio of 5 v / v%, 10 v / v%, and 25 v / v% is used, a trimer type is formed on the glass substrate. The residue of fine crystals of acetone peroxide was not confirmed. That is, after the wiping treatment, a slight film layer of the solution remains on the glass substrate, but when the solvent in the remaining film layer of the solution evaporates, a fine crystalline residue of trimeric acetone peroxide is left. It was not found.

  On the other hand, when a 1: 1 (v / v) mixed solvent of methanol and acetonitrile and a (methanol / acetonitrile) / water mixed solution with a mixing ratio of 50 v / v% are used, after wiping treatment, on the glass substrate In addition, although a film layer of the solution remained slightly, when the solvent in the film layer of the remaining solution was evaporated, a residue of fine crystals of trimeric acetone peroxide was found.

(Condition 3)
Wiping is performed using Seprafilm (registered trademark) of the base material that has not been subjected to the above-described monoclonal antibody immobilization treatment.

  The base film was absorbed with 2 ml of an acetonitrile / water mixture dissolved in pure water at a concentration of 50 v / v% to make it infiltrated, and then the trimeric acetone peroxide microparticles on the glass substrate were absorbed. Cover the crystal powder. In this state, a total of 0.5 mg of trimeric acetone peroxide microcrystal powder was dissolved by an infiltrating acetonitrile / water mixture, and then the substrate film was removed.

  After finishing the wiping process, the surface of the glass substrate is observed.

  In the range observed visually, the residue of fine crystals of trimeric acetone peroxide was confirmed on the glass substrate. That is, when the film of the base material is removed, a film layer of the solution slightly remains on the glass substrate, but the solvent in the film layer of the remaining solution evaporates, and the finer form of trimeric acetone peroxide is removed. A crystalline residue was found.

  In the above-mentioned conditions 1 to 3, when a wiping treatment material or a film of the base material is removed, a phenomenon that a film layer of a liquid slightly remains on the glass substrate has been found similarly.

  Of course, in condition 3, the film layer of the remaining liquid is a solution in which trimeric acetone peroxide is dissolved at a concentration of 0.5 mg / 2 ml in an acetonitrile / water mixture.

  Further, when acetonitrile (100 v / v%) is used in condition 1, every time in condition 2, a 1: 1 (v / v) mixed solvent of methanol and acetonitrile, the mixing ratio is 50 v / v%. When the (methanol / acetonitrile) / water mixed solution is used, the film layer of the slightly remaining liquid is a solution in which trimeric acetone peroxide is dissolved in an appropriate concentration in each mixed solvent. It is judged that. The concentration is considerably reduced compared with the initial concentration of 0.5 mg / 2 ml, but when the solvent in the remaining film layer of the solution evaporates, the trimeric acetone peroxide has a fine concentration. The concentration is sufficient to produce a crystalline residue.

  On the other hand, in the condition 1, when the acetonitrile / water mixture having acetonitrile concentrations of 5 v / v%, 10 v / v%, 25 v / v%, 50 v / v% is used, and in the condition 2, the mixing ratio is When a 5 v / v%, 10 v / v%, or 25 v / v% (methanol / acetonitrile) / water mixture is used, the film layer of the slightly remaining liquid substantially contains trimeric acetone peroxide. It is judged that the liquid does not contain it.

  Therefore, by using the above-mentioned wiping treatment material in which the monoclonal antibody is immobilized using the acetonitrile / water mixture (methanol / acetonitrile) / water mixture having the above mixing ratio, It was verified that the fine crystalline powder of acetone-type acetone was wiped off efficiently.

  A treatment material for a peroxide derivative-type explosive according to the present invention and a removal method using the treatment material include a peroxide derivative-type explosive to be treated, for example, a trimeric acetone peroxide (TATP). ) Can be conveniently used for the purpose of removing it easily and safely.

1. 1. Water-absorbing material 2. Monoclonal antibody Water absorbing material 4. 4. Water-absorbing material holding antibodies Acetone peroxide 6. 6. Glass substrate Pipette 8. Solvent 9. Linker 10. Polyvinyl alcohol

(Trust number)
As a hybridoma cell producing a monoclonal antibody having a binding ability to the trimeric acetone peroxide (TATP) represented by the formula (I) used in the present invention,
Hybridoma cell line: NECP-C57Z 3B-7E is based on the Budapest Treaty. National Institute of Advanced Industrial Science and Technology, Patent Biological Deposit Center In addition, an international deposit (May 12, 2009) has been made.

Claims (27)

Peroxide derivative type explosive, acetone peroxide of the following formula (I) (TATP: 3,3,6,6,9,9-hexamethyl-1,2,4,5,7,6-hexaoxacyclononane ) By wiping and removing the solid powder,

The method of wiping and removing is as follows:
Dissolving a solid powder of acetone peroxide of formula (I) in an appropriate amount of a solvent capable of dissolving the acetone peroxide of formula (I);
Using a wiping treatment material made of a liquid-absorbing material having a liquid-absorbing property with respect to the solvent, the wiping treatment material absorbs the liquid in which acetone peroxide of the formula (I) is dissolved, and is wiped off. It has a process,
The wiping treatment material is
A monoclonal antibody having binding ability to acetone peroxide of formula (I) is immobilized in the liquid-absorbing material;
The acetone peroxide of formula (I) dissolved in the solution of acetone peroxide of formula (I) absorbed in the wiping treatment material is immobilized by the antigen-antibody reaction. A method for wiping off explosives, comprising forming a complex with an antibody. The monoclonal antibody having binding ability to acetone peroxide of formula (I) is:
A monoclonal antibody against a low molecular weight compound having a structure similar to the characteristic structure of acetone peroxide of the formula (I), and cross-reactive with the peroxide peroxide of the formula (I) The explosive wiping and removing method according to claim 1, comprising: The low molecular weight compound having a structure similar to the characteristic structure of acetone peroxide of the formula (I) is a dicarboxylic acid compound having a structure represented by the following formula (II): 3- [12- (2 -Carboxyethyl) -9,12-dimethyl-7,8,10,11,13,14-hexaoxa-spiro- [5.8] tetradec-9-yl] -propionic acid (3- [12- (2-carboxyethyl) -9,12-dimethyl-7,8,10,11,13,14-hexaoxa-spiro- [5.8] tetradec-9-yl] -propanoic acid)

The method for wiping and removing explosives according to claim 2. The monoclonal antibody having binding ability to acetone peroxide of the formula (I) is:
A non-human mammal can be obtained by using a modified protein obtained by binding a low molecular weight compound having a structure similar to the characteristic structure of acetone peroxide of formula (I) on a carrier protein as an immunogen. A monoclonal antibody against the low molecular weight compound created by immunization,
4. The method for wiping off explosives according to claim 2 or 3, wherein the antibody has cross-reactivity with acetone peroxide of formula (I). The explosive wiping and removing method according to claim 4, wherein the mammal other than a human is a mouse. 6. The modified protein obtained by binding the low molecular compound on a carrier protein, wherein the carrier protein is selected from keyhole limpet hemocyanin (Keyhole Limpet Hemocyanin). The explosive wiping off method described. A compound having a carboxyl group (—COOH) in the molecule is selected as a low molecular weight compound having a structure similar to the characteristic structure of acetone peroxide of formula (I),
A modified protein obtained by binding a compound having a carboxyl group (—COOH) in the molecule onto a carrier protein comprises the carboxyl group (—COOH) and an amino group (—NH ₂ ) on the carrier protein. A compound having a carboxyl group (—COOH) in the molecule is bonded through an amide bond (—CO—NH—) between the two groups. The explosive wiping off method described in the item. Formation of an amide bond (—CO—NH—) between the carboxyl group (—COOH) and an amino group (—NH ₂ ) on the carrier protein is performed using a carbodiimide method. The method for wiping and removing explosives according to claim 7. As a monoclonal antibody immobilized in the liquid absorbing material,
The monoclonal antibody against the dicarboxylic acid compound having the structure represented by the formula (II) produced by a hybridoma cell line: NECP-C57Z 3B-7E (FERM BP-11125) is used. How to wipe off explosives. The method for wiping and removing explosives according to any one of claims 1 to 9, wherein the monoclonal antibody is immobilized in the liquid-absorbing material with polyvinyl alcohol. The method for wiping and removing explosives according to any one of claims 1 to 9, wherein the monoclonal antibody is immobilized in the liquid-absorbing material through a linker composed of at least a protein. . 12. The explosive wiping and removing method according to claim 11, wherein the at least protein linker is at least one of protein A and protein G. The method for wiping and removing explosives according to any one of claims 1 to 12, wherein the wiping treatment material is used in a step of wiping and removing after at least water or the solvent is absorbed. In the operation of dissolving the solid peroxide of acetone peroxide of the formula (I) in an appropriate amount of a solvent capable of dissolving the acetone peroxide of the formula (I),
The solid powder of acetone peroxide of formula (I) is dissolved using an appropriate amount of a solvent capable of dissolving acetone peroxide of formula (I) to form a solution. 14. The explosive wiping and removing method according to any one of items 13 to 13. The solvent capable of dissolving acetone peroxide of formula (I) includes at least one organic solvent selected from the group of organic solvents consisting of acetonitrile, methanol, ethanol, acetone, ethanolamine, and methyl ethyl ketone. The explosive wiping removal method as described in any one of Claims 1-14. Peroxide derivative type explosive, acetone peroxide of the following formula (I) (TATP: 3,3,6,6,9,9-hexamethyl-1,2,4,5,7,6-hexaoxacyclononane ) A wiping treatment material usable for wiping and removing solid powder

The wiping treatment material is
Made of a liquid-absorbing material having a liquid-absorbing property to a solvent capable of dissolving acetone peroxide of the formula (I),
An explosive wiping treatment material, wherein a monoclonal antibody having binding ability to acetone peroxide of formula (I) is immobilized in the liquid absorbing material. The monoclonal antibody having binding ability to acetone peroxide of formula (I) is:
A monoclonal antibody against a low molecular weight compound having a structure similar to the characteristic structure of acetone peroxide of the formula (I), and cross-reactive with the peroxide peroxide of the formula (I) The explosive wiping treatment material according to claim 16, comprising: The low molecular weight compound having a structure similar to the characteristic structure of acetone peroxide of the formula (I) is a dicarboxylic acid compound having a structure represented by the following formula (II): 3- [12- (2 -Carboxyethyl) -9,12-dimethyl-7,8,10,11,13,14-hexaoxa-spiro- [5.8] tetradec-9-yl] -propionic acid (3- [12- (2-carboxyethyl) -9,12-dimethyl-7,8,10,11,13,14-hexaoxa-spiro- [5.8] tetradec-9-yl] -propanoic acid)

The explosive wiping treatment material according to claim 17, wherein The monoclonal antibody having binding ability to acetone peroxide of the formula (I) is:
A non-human mammal can be obtained by using a modified protein obtained by binding a low molecular weight compound having a structure similar to the characteristic structure of acetone peroxide of formula (I) on a carrier protein as an immunogen. A monoclonal antibody against the low molecular weight compound created by immunization,
19. The explosive wiping treatment material according to claim 17 or 18, characterized in that the antibody has cross-reactivity with acetone peroxide of formula (I). The explosive wiping treatment material according to claim 19, wherein the non-human mammal is a mouse. 21. The modified protein obtained by binding the low molecular weight compound on a carrier protein, wherein keyhole limpet hemocyanin is selected as the carrier protein. The explosive wiping treatment material described. A compound having a carboxyl group (—COOH) in the molecule is selected as a low molecular weight compound having a structure similar to the characteristic structure of acetone peroxide of formula (I),
A modified protein obtained by binding a compound having a carboxyl group (—COOH) in the molecule onto a carrier protein comprises the carboxyl group (—COOH) and an amino group (—NH ₂ ) on the carrier protein. The compound having a carboxyl group (—COOH) in the molecule is bonded through an amide bond (—CO—NH—) between the two groups. Explosive wiping treatment material as described in the paragraph. Formation of an amide bond (—CO—NH—) between the carboxyl group (—COOH) and an amino group (—NH ₂ ) on the carrier protein is performed using a carbodiimide method. The explosive wiping treatment material according to claim 22. As a monoclonal antibody immobilized in the liquid absorbing material,
The monoclonal antibody against the dicarboxylic acid compound having the structure represented by the formula (II) produced by a hybridoma cell line: NECP-C57Z 3B-7E (FERM BP-11125) is used. Explosive wiping treatment material. The explosive wiping treatment material according to any one of claims 16 to 24, wherein the monoclonal antibody is immobilized in the liquid-absorbing material with polyvinyl alcohol. The explosive wiping treatment material according to any one of claims 16 to 24, wherein the monoclonal antibody is immobilized in the liquid-absorbing material through at least a linker made of a protein. . 27. The explosive wiping treatment material according to claim 26, wherein the at least protein linker is at least one of protein A and protein G.

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