TWI900607B - Composition containing tarc and method for enhancing storage stability of tarc - Google Patents

Abstract

The subject of the present invention is to provide a composition containing TARC with higher storage stability. The present invention can solve the above problem by a composition containing TARC (Thymus and activation-regulated chemokine), which includes TARC and at least one surfactant selected from the group consisting of cationic surfactants and anionic surfactants, and is in liquid form.

Description

Composition containing TARC and method for improving storage stability of TARC

The present invention relates to a composition containing TARC. More specifically, the present invention relates to a composition capable of stably preserving TARC for a long period of time. Furthermore, the present invention relates to a method and kit for measuring TARC, a method for improving the storage stability of TARC, and an agent for preventing TARC adsorption.

Thymus and activation-regulated chemokine (TARC), a C-C chemokine ligand 17 (CCL17), is a chemokine that promotes leukocyte migration. TARC attracts Th2 cells, a type of lymphocyte, to lesions, leading to IgE production and eosinophil infiltration and activation. This exacerbates allergic reactions and, in turn, exacerbates the symptoms of atopic dermatitis (Non-Patent Document 1).

For atopic dermatitis, rapid and reliable sedation of inflammation is crucial. Compared to other indicators of atopic dermatitis severity, such as serum IgE, peripheral blood eosinophil count, and serum LDH, TARC has a better correlation with atopic dermatitis severity and more sensitively reflects the condition (Non-Patent Document 2). Therefore, using TARC as a biomarker allows for objective and rapid assessment of atopic dermatitis severity and effectiveness when selecting or changing atopic dermatitis treatments.

To quantify the target component in a biological sample, a calibration sample is required. A calibration sample is a sample containing the target component and is used as an internal standard or concentration calibration standard (calibrant). To obtain accurate quantitative values, a calibration sample must be stable over time or temperature. For ease of handling, a flowable solution (sometimes referred to as a "liquid" below) is preferred. In this case, ideal characteristics for calibration samples include maintaining biological activity (e.g., antigenicity for specific antibodies; binding activity for specific binding targets of antibodies or lectins; physiological and enzymatic activity of peptide hormones; and the stereostructure of proteins supporting these activities), preventing adsorption to containers, and maintaining preservative properties.

Known methods for preserving calibration samples intended for use in immunological assays in liquid form include: stabilizing antigens by coexisting the calibration sample with casein and/or whey protein (Patent Document 1), stabilizing insulin by coexisting the calibration sample with a bile acid amide derivative (Patent Document 2), and stabilizing soluble interleukin-2 receptor (sIL-2R) by coexisting the calibration sample with a chelating agent (Patent Document 3). While stabilization methods must be developed based on the properties of the substance being stabilized, detailed studies on TARC liquid calibration samples have not yet been conducted.

[Prior Art Literature] [Patent Literature]

[Patent Document 1] Japanese Patent Application Laid-Open No. 08-005634

[Patent Document 2] Japanese Patent Application Laid-Open No. 08-012593

[Patent Document 3] Japanese Patent Application Laid-Open No. 2010-230660

[Non-patent literature]

[Non-patent document 1] J Allergy Clin Immunol 107: 535-541, 2001

[Non-patent document 2] Journal of the Japanese Dermatological Society 116(1): 27-39, 2006

The object of the present invention is to provide a composition containing TARC with higher storage stability.

The inventors attempted to produce a TARC-containing composition with enhanced storage stability. Subsequently, they discovered that the stability of TARC is enhanced when TARC is added to a solution containing at least one surfactant selected from the group consisting of cationic and anionic surfactants. The present invention is based on this finding.

Specifically, the present invention is as follows.

<1> A composition containing TARC (thymic activation regulatory factor), which comprises TARC and at least one surfactant selected from the group consisting of cationic surfactants and anionic surfactants, and is in liquid form.

<2> The composition described in <1> is used as a calibration sample solution for the determination of TARC.

<3> The composition described in <1> or <2> is filled in a storage container.

<4> The composition described in <3>, wherein the storage container is plastic or glass.

<5> The composition according to any one of <1> to <4>, wherein the concentration of TARC relative to the composition is 10 pg/mL to 1 μg/mL.

<6> The composition according to any one of <1> to <5>, wherein the concentration of the surfactant is 0.00001% by mass to 1% by mass relative to the composition.

<7> The composition according to any one of <1> to <6>, wherein the cationic surfactant is an alkylamine salt type or a quaternary ammonium salt type cationic surfactant, and the anionic surfactant is a cholic acid type, sulfate type, carboxylic acid type, or sulfonic acid type anionic surfactant.

<8> The composition according to any one of <1> to <7>, wherein, when the TARC concentration relative to the composition is 500 pg/mL, the residual rate of TARC after storage at 37°C in a plastic container for 28 days is 80% or more.

<9> The composition according to any one of <1> to <8>, wherein, when the TARC concentration relative to the composition is 500 pg/mL, the residual rate of TARC after storage at 4°C in a plastic container for 28 days is 90% or greater.

<10> The composition according to any one of <1> to <9>, wherein, when the concentration of TARC in the composition is 2000 pg/mL, the residual rate of TARC after storage in a glass container at 10°C for 6 hours is 55% or more.

<11> The composition according to any one of <1> to <10>, wherein the cationic surfactant is lauryltrimethylammonium chloride or cocamine acetate, and the anionic surfactant is sodium cholate, sodium lauryl sulfate, sodium lauryl methylalanine, or sodium β-naphthalenesulfonate formalin condensate.

<12> A method for determining TARC using the composition described in any one of <1> to <11>.

<13> A TARC assay kit comprising the composition described in any one of <1> to <11>.

<14> A method for improving the storage stability of TARC, comprising the step of contacting the TARC with a solution containing at least one surfactant selected from the group consisting of cationic surfactants and anionic surfactants.

<15> The method for improving the storage stability of TARC as described in <14>, comprising the step of adjusting the concentration of the TARC in the solution to be between 10 pg/mL and 1 μg/mL.

<16> The method for improving the storage stability of TARC as described in <14> or <15>, wherein the concentration of the surfactant is 0.00001 mass% to 1 mass% relative to the solution.

<17> The method for improving the storage stability of TARC according to any one of <14> to <16>, wherein the cationic surfactant is an alkylamine salt type or a quaternary ammonium salt type cationic surfactant, and the anionic surfactant is a cholic acid type, sulfate type, carboxylic acid type, or sulfonic acid type anionic surfactant.

<18> The method for improving the storage stability of TARC according to any one of <14> to <17>, wherein the cationic surfactant is lauryltrimethylammonium chloride or cocamide acetate, and the anionic surfactant is sodium cholate, sodium lauryl sulfate, sodium lauryl methylalanine, or sodium β-naphthalenesulfonate formalin condensate.

<19> An adsorption inhibitor for preventing TARC in a solution containing TARC from being adsorbed on a container, comprising as an active ingredient at least one surfactant selected from the group consisting of cationic surfactants and anionic surfactants.

<20> The adsorption inhibitor according to <19>, wherein the concentration of TARC in the solution containing TARC is 10 pg/mL to 1 μg/mL relative to the solution.

<21> The adsorption inhibitor described in <19> or <20> is used in a manner such that the concentration of the surfactant is 0.00001% by mass to 1% by mass relative to the solution.

<22> The adsorption inhibitor according to any one of <19> to <21>, wherein the cationic surfactant is an alkylamine salt type or a quaternary ammonium salt type cationic surfactant, and the anionic surfactant is a cholic acid type, sulfate type, carboxylic acid type, or sulfonic acid type anionic surfactant.

<23> The adsorption inhibitor according to any one of <19> to <22>, wherein the cationic surfactant is lauryltrimethylammonium chloride or cocamide acetate, and the anionic surfactant is sodium cholate, sodium lauryl sulfate, sodium lauryl methylalanine, or sodium β-naphthalenesulfonate formalin condensate.

The present invention provides a TARC-containing composition with high storage stability, particularly a TARC-containing calibration sample. Therefore, the present invention allows for accurate quantification of TARC in biological samples, enabling precise assessment of the severity of atopic dermatitis.

(TARC)

In this specification, "TARC" refers to thymic activation-regulated trend factor (CCL17). TARC is a trend factor that regulates leukocyte migration. TARC functions by attracting Th2 cells, a type of lymphocyte, to lesions, leading to IgE production and eosinophil infiltration and activation. Using TARC as a biomarker allows for objective and rapid assessment of atopic dermatitis severity when selecting or changing treatments.

TARC can be measured using known methods, such as immunological methods. Examples of immunological methods include ELISA, enzyme immunoassay, surface plasmon resonance, latex agglutination immunoassay (LTIA), chemiluminescent immunoassay, electrochemical luminescent immunoassay, fluorescent antibody method, radioimmunoassay, Western blotting, immunochromatography, and high-performance liquid chromatography (HPLC).

The TARC contained in the composition of the present invention may be commercially available or produced or purified. The TARC contained in the composition of the present invention may be produced in vitro or extracted from organisms.

(TARC concentration)

Considering the stability of TARC, the concentration of TARC in the composition of the present invention is preferably 10 pg/mL to 1 μg/mL, more preferably 50 pg/mL to 500 ng/mL, further preferably 100 pg/mL to 100 ng/mL, and most preferably 100 pg/mL to 50 ng/mL, but is not limited to these.

(Cationic surfactants and anionic surfactants)

In this specification, anionic surfactants refer to surfactants whose hydrophobic groups partially dissociate into negative (minus) ions when dissolved in water. In this specification, surfactants with hydrophilic groups that do not ionize when dissolved in water are referred to as nonionic surfactants, surfactants whose hydrophobic groups partially dissociate into positive (plus) ions when dissolved in water are referred to as cationic surfactants, and surfactants that exhibit the properties of anionic surfactants in alkaline regions and cationic surfactants in acidic regions are referred to as amphoteric surfactants.

Cationic surfactants are preferably alkylamine salts and quaternary ammonium salts. Anionic surfactants are preferably cholic acid, sulfate (sulfate) esters, carboxylic acids, and sulfonic acids.

Examples of cationic surfactants in the alkylamine salt type include salts of amines containing 1 to 3 alkyl groups with 8 to 22 carbon atoms, such as monododecylamine, monooctadecylamine, dioctadecylamine, and trioctadecylamine, and inorganic acids such as hydrochloric acid and sulfuric acid, or lower carboxylic acids such as acetic acid, lactic acid, and citric acid. Specific examples include laurylamine hydrochloride [CAS No.: 929-73-7, manufactured by Tokyo Chemical Industry Co., Ltd.], cocoamine acetate [CAS No.: 61790-57-6, product name: Acetamin 24, manufactured by Kao Corporation], and stearylamine acetate [CAS No.: 2190-04-7, product name: Acetamin 86, manufactured by Kao Corporation].

Examples of quaternary ammonium salts include ammonium salts containing 1 to 3 alkyl groups with 8 to 22 carbon atoms, such as dodecyltrimethylammonium, octadecyltrimethylammonium, hexadecyltrimethylammonium, didecyldimethylammonium, and benzylmethyltetradecylammonium, and chlorine, bromine, or the like. Specifically, for example, lauryltrimethylammonium chloride [CAS No.: 112-00-5, product name: Quartamin (registered trademark) 24P, manufactured by Kao Corporation], dodecyltrimethylammonium bromide [CAS No.: 1119-94-4, manufactured by Tokyo Chemical Industry Co., Ltd.], octadecyltrimethylammonium chloride [CAS No.: 112-03-8, manufactured by Tokyo Chemical Industry Co., Ltd.], and distearyldimethylammonium chloride [CAS No.: 107-64-2, product name: Quartamin (registered trademark) D86P, manufactured by Kao Corporation] can be cited.

Examples of anionic surfactants of the cholic acid type include sodium deoxycholate [CAS No.: 302-95-4, manufactured by FUJIFILM Wako Co., Ltd.] and sodium cholate [CAS No.: 361-09-1, manufactured by FUJIFILM Wako Co., Ltd.].

Examples of sulfate esters include higher alcohol sulfates such as lauryl sulfate, and salts of polyoxyethylene alkyl ether sulfates such as polyoxyethylene lauryl ether sulfate with sodium, ammonium, and the like. Specific examples include higher alcohol sulfates such as sodium lauryl sulfate [CAS No. 151-21-3, manufactured by Sigma-Aldrich] and ammonium lauryl sulfate [CAS No. 2235-54-3, Latemul (registered trademark) AD-25, manufactured by Kao Corporation]; and polyoxyethylene alkyl ether sulfates such as sodium polyoxyethylene lauryl ether sulfate [CAS No. 68585-34-2, product name: Emal 20C, manufactured by Kao Corporation].

Examples of the carboxylic acid type include higher fatty acids with 8 to 22 carbon atoms, such as dodecanoic acid (lauric acid), tetradecanoic acid (myristic acid), hexadecanoic acid (palmitic acid), octadecanoic acid (stearic acid), and cis-9-octadecenoic acid (oleic acid), and salts of alkaline metals such as sodium and potassium. Specifically, for example, sodium laurate [CAS No.: 629-25-4, manufactured by FUJIFILM Wako Co., Ltd.], sodium myristate [CAS No.: 822-12-8, manufactured by FUJIFILM Wako Co., Ltd.], sodium palmitate [CAS No.: 408-35-5, manufactured by Tokyo Chemical Industry Co., Ltd.], sodium lauroylsarcosinate [CAS No.: 137-16-6, manufactured by Tokyo Chemical Industry Co., Ltd.], sodium polyoxyethylene lauryl ether acetate [CAS No.: 33939-64-9, product name: Enagicol EC-30, manufactured by Lion Specialty Chemicals Co., Ltd.], and sodium lauroyl methylalanine [CAS No.: 21539-58-2, product name: Enagicol L-30AN, manufactured by Lion Specialty Chemicals Co., Ltd.] etc.

Examples of sulfonic acid-type sulfonic acids include alkylbenzenesulfonic acids such as dodecylbenzenesulfonic acid, naphthalenesulfonic acids such as dibutylnaphthalenesulfonic acid, formalin condensates of naphthalenesulfonic acid, and salts of sulfosuccinic acids such as dioctylsulfosuccinic acid with sodium. Specific examples include sodium laurylbenzenesulfonate [CAS No.: 25155-30-0, manufactured by FUJIFILM Wako Co., Ltd.], β-naphthalenesulfonic acid formalin condensate sodium salt [CAS No.: 9084-06-4, product name: Demol (registered trademark) RN, manufactured by Kao Corporation], and sodium dioctylsulfosuccinate [CAS No.: 577-11-7, product name: Neocol (registered trademark) SW, manufactured by Daiichi Industry Co., Ltd.].

These anionic or cationic surfactants are sold by various cleaning agent or reagent manufacturers and can be purchased for use. Furthermore, these anionic or cationic surfactants can be used alone or in combination of two or more.

The cationic or anionic surfactant used in the composition of the present invention is preferably cocamide acetate, lauryltrimethylammonium chloride, sodium cholate, sodium lauryl sulfate, sodium lauryl methylalanine, or sodium β-naphthalenesulfonate formalin condensate. In particular, for plastic containers, lauryltrimethylammonium chloride, sodium cholate, sodium lauryl sulfate, or sodium lauryl methylalanine are more preferably used. For glass containers, lauryltrimethylammonium chloride, cocamide acetate, or sodium β-naphthalenesulfonate formalin condensate are more preferably used.

While other types of surfactants may be used in addition to anionic or cationic surfactants as long as they do not impair the effects of the present invention, it is preferred that they not be used. Furthermore, anionic and cationic surfactants may be used together, but it is preferred that only one of them be used.

In the composition of the present invention, the order of adding the cationic surfactant or anionic surfactant and TARC is not particularly limited, as long as the effects of the present invention can be achieved.

(Concentration of surfactant)

In the composition of the present invention, the concentration of the cationic surfactant or anionic surfactant is preferably 0.00001% to 1% by mass relative to the composition, more preferably 0.0001% to 1% by mass, further preferably 0.001% to 0.5% by mass, and most preferably 0.001% to 0.1% by mass, based on the stability of TARC. However, the concentration is not limited to these.

(Storage container)

The composition of the present invention is preferably filled in a storage container. The material of the storage container is not particularly limited, as long as it can achieve the effects of the present invention and is sealable. At least part or all of the portion in contact with the composition can be plastic [for example, olefin resins, styrene resins, acrylic resins, polyester resins, polycarbonate resins, fluororesins, chlorine-containing resins (such as polyvinyl chloride), polyamide resins, polyacetal resins, polyphenylene ether resins (modified polyphenylene ether), polyarylate, polysulfone, polyimide resins, cellulose resins (such as cellulose acetate), hydrocarbon resins (including halogen-substituted products), etc.], metal (such as aluminum), glass, etc. From the perspective of manufacturing, transporting, and storing calibration samples, plastic or glass is preferred. Among plastics, olefin resins are preferred, and polypropylene is even more preferred.

The storage container may be made of a single material or a combination of two or more materials, preferably a single material. The storage container may include a container body and a lid. In this case, the container body and lid may be made of different materials. Furthermore, the storage container, particularly the container body, is preferably transparent enough to allow the liquid contents to be visible from the outside.

The storage container can be either rigid or flexible, and examples include ampoules, vials, soft bags, injection containers, and glass bottles. From the perspectives of ease of use and TARC stability, the storage container is preferably in the form of a plastic eye dropper, comprising a container body and a lid, particularly a cylindrical eye dropper. Furthermore, from the perspectives of ease of use and TARC stability, the storage container is preferably a glass ampoule, or a glass container body and a lid, particularly a cylindrical glass container and a rubber lid.

(Composition)

The composition of the present invention can be used as a calibration sample solution in TARC measurements. In this specification, a calibration sample solution refers to a sample solution containing the substance being measured at a certain concentration, used for accurate measurement of the substance being measured. This includes standards, calibrants, controls, and internal standards. Examples of the composition of the present invention include a pre-prepared solution of TARC mixed with an anionic or cationic surfactant in a solvent.

Furthermore, in the TARC measurement method, "using a composition" means using a composition for accurate TARC measurement. For example, it means using a liquid composition containing TARC as a calibration sample solution (standard substance, calibrator, control substance, internal standard substance, etc.).

The pH of the composition of the present invention is, for example, 4.0-9.5, 5.0-9.0, 6.0-8.5, 6.5-8.0, or 7.0-8.0.

The pH can be adjusted using pH adjustment reagents known to the industry, such as sodium hydroxide or hydrochloric acid.

The composition of the present invention is not particularly limited, provided it does not impair the effectiveness of the present invention. If TARC is measured by immunological assays, the effectiveness of the present invention is not impaired, and the composition does not interfere with all or part of the reactions that comprise the assay system, such as the antigen-antibody reaction, the labeling reaction for detecting biotin-avidin, and the enzyme reaction. Depending on the intended purpose, various components commonly used in immunological assays can be appropriately selected. Examples include buffers such as acetic acid, citric acid, phosphoric acid, PBS (phosphate-buffered saline), HEPES, MES, Tris, glycine, boric acid, carbonic acid, or Good's buffer; components that promote antigen-antibody reactions (polymers such as polyethylene glycol and polyvinylpyrrolidone); glycoproteins or peptides (such as BSA and casein); amino acids; salts (such as sodium chloride and potassium chloride); carbohydrates (such as sucrose and cyclodextrin); and preservatives (such as sodium azide and ProClin 300). PBS with a pH of 6.5 to 8.0 is preferred.

In this specification, "improved storage stability" or "improved storage stability" means that the majority of the TARC contained in a TARC-containing solution remains in a state without decomposition, structural change, or adsorption to the container over a prolonged period of time. Therefore, there is no significant difference between the initial TARC value in the solution and the value measured after storage.

More specifically, "improved storage stability" or "improved storage stability" may mean, for example, that at least 80% of the TARC contained in a solution containing TARC at a concentration of 10 pg/mL to 1 μg/mL remains undecomposed, structurally altered, or adsorbed to the container over a 28-day period at 37°C. Therefore, the TARC content of the solution measured after 28 days of storage at 37°C in a plastic container is at least 80% of the initial value.

Furthermore, "improved storage stability" or "improved storage stability" may mean, for example, that 90% or more of the TARC contained in a solution containing TARC at a concentration of 10 pg/mL to 1 μg/mL remains undecomposed, structurally altered, or adsorbed to the container for 28 days at 4°C. Therefore, the TARC value measured in the solution after 28 days of storage at 4°C in a plastic container is at least 90% of the initial value.

Furthermore, "improved storage stability" or "improved storage stability" may mean, for example, that 55% or more of the TARC contained in a solution containing TARC at a concentration of 10 pg/mL to 1 μg/mL remains undecomposed, structurally altered, or adsorbed to the container within 6 hours at 10°C. Therefore, the TARC value measured in the solution after storage at 10°C in a glass container for 6 hours is at least 55% of the initial value.

(Biological samples used for TARC determination)

There are no particular limitations on the biological sample used for TARC measurement; any sample capable of TARC measurement is preferred. Blood, serum, or plasma is preferred. The biological sample may be pretreated as needed. Preferably, the biological sample is collected from a human.

(TARC test kit)

The TARC assay kit of the present invention uses the composition of the present invention to conveniently and accurately measure TARC. Examples of TARC assay kits include those using immunological methods. The TARC assay kit of the present invention may include reagents for measuring the concentration of TARC in the human body using immunological methods. Examples of immunological methods include ELISA, enzyme immunoassay, surface plasmon resonance, latex agglutination immunoassay (LTIA), chemiluminescent immunoassay, electrochemical luminescent immunoassay, fluorescent antibody method, radioimmunoassay, Western blotting, immunochromatography, and high-performance liquid chromatography (HPLC).

The TARC assay kit of the present invention can be used to assess the severity of atopic dermatitis when selecting treatments or medications for the condition and evaluating treatment effectiveness.

In addition, the TARC assay kit of the present invention may also include instructions for use. The TARC assay kit may also include optional components, such as a buffer, stabilizer, sample diluent, pH adjuster, reaction vessel, etc.

(Methods to improve TARC's storage stability)

The method for improving the storage stability of TARC of the present invention includes contacting TARC with a solution containing a cationic surfactant or an anionic surfactant. TARC can be added after the cationic surfactant or anionic surfactant is added to the solution, or the cationic surfactant or anionic surfactant can be added after the TARC is added to the solution. The solution is preferably a buffer such as PBS, HEPES, MES, CHES, or Tris. PBS with a pH of 6.5 to 8.0 is more preferred.

(TARC concentration)

In the method for improving the storage stability of TARC of the present invention, the concentration of TARC in the solution is preferably 10 pg/mL to 1 μg/mL, more preferably 50 pg/mL to 500 ng/mL, further preferably 100 pg/mL to 100 ng/mL, and most preferably 100 pg/mL to 50 ng/mL, considering TARC stability. However, the concentration is not limited to these.

Cationic surfactants are preferably alkylamine salts and quaternary ammonium salts. Anionic surfactants are preferably cholic acid, sulfate (sulfate) esters, carboxylic acids, and sulfonic acids.

Examples of cationic surfactants in the alkylamine salt type include salts of amines containing 1 to 3 alkyl groups with 8 to 22 carbon atoms, such as monododecylamine, monooctadecylamine, dioctadecylamine, and trioctadecylamine, and inorganic acids such as hydrochloric acid and sulfuric acid, or lower carboxylic acids such as acetic acid, lactic acid, and citric acid. Specific examples include laurylamine hydrochloride [CAS No.: 929-73-7, manufactured by Tokyo Chemical Industry Co., Ltd.], cocoamine acetate [CAS No.: 61790-57-6, product name: Acetamin 24, manufactured by Kao Corporation], and stearylamine acetate [CAS No.: 2190-04-7, product name: Acetamin 86, manufactured by Kao Corporation].

Examples of quaternary ammonium salts include ammonium salts containing 1 to 3 alkyl groups with 8 to 22 carbon atoms, such as dodecyltrimethylammonium, octadecyltrimethylammonium, hexadecyltrimethylammonium, didecyldimethylammonium, and benzylmethyltetradecylammonium, and chlorine, bromine, or the like. Specifically, for example, lauryltrimethylammonium chloride [CAS No.: 112-00-5, product name: Quartamin (registered trademark) 24P, manufactured by Kao Corporation], dodecyltrimethylammonium bromide [CAS No.: 1119-94-4, manufactured by Tokyo Chemical Industry Co., Ltd.], octadecyltrimethylammonium chloride [CAS No.: 112-03-8, manufactured by Tokyo Chemical Industry Co., Ltd.], and distearyldimethylammonium chloride [CAS No.: 107-64-2, product name: Quartamin (registered trademark) D86P, manufactured by Kao Corporation] can be cited.

Examples of bile acid-based anionic surfactants include sodium deoxycholate [CAS No.: 302-95-4, manufactured by FUJIFILM Wako Co., Ltd.] and sodium cholate [CAS No.: 361-09-1, manufactured by FUJIFILM Wako Co., Ltd.].

Examples of sulfate esters include higher alcohol sulfates such as lauryl sulfate, and salts of polyoxyethylene alkyl ether sulfates such as polyoxyethylene lauryl ether sulfate with sodium, ammonium, and the like. Specifically, examples include higher alcohol sulfates such as sodium lauryl sulfate [CAS No. 151-21-3, manufactured by Sigma-Aldrich] and ammonium lauryl sulfate [CAS No. 2235-54-3, Latemul (registered trademark) AD-25, manufactured by Kao Corporation]; and polyoxyethylene alkyl ether sulfates such as sodium polyoxyethylene lauryl ether sulfate [CAS No. 68585-34-2, product name: Emal 20C, manufactured by Kao Corporation].

Examples of the carboxylic acid type include higher fatty acids with 8 to 22 carbon atoms, such as dodecanoic acid (lauric acid), tetradecanoic acid (myristic acid), hexadecanoic acid (palmitic acid), octadecanoic acid (stearic acid), and cis-9-octadecenoic acid (oleic acid), and salts of alkaline metals such as sodium and potassium. Specifically, for example, sodium laurate [CAS No.: 629-25-4, manufactured by FUJIFILM Wako Co., Ltd.], sodium myristate [CAS No.: 822-12-8, manufactured by FUJIFILM Wako Co., Ltd.], sodium palmitate [CAS No.: 408-35-5, manufactured by Tokyo Chemical Industry Co., Ltd.], sodium lauryl sarcosine [CAS No.: 137-16-6, manufactured by Tokyo Chemical Industry Co., Ltd.], sodium polyoxyethylene lauryl ether acetate [CAS No.: 33939-64-9, product name: Enagicol EC-30, manufactured by Lion Specialty Chemicals Co., Ltd.], and sodium lauryl methylalanine [CAS No.: 21539-58-2, product name: Enagicol L-30AN, manufactured by Lion Specialty Chemicals Co., Ltd.] can be mentioned. Chemicals Co., Ltd.] etc.

Examples of sulfonic acid-type sulfonic acids include alkylbenzenesulfonic acids such as dodecylbenzenesulfonic acid, naphthalenesulfonic acids such as dibutylnaphthalenesulfonic acid, formalin condensates of naphthalenesulfonic acid, and salts of sulfosuccinic acids such as dioctylsulfosuccinic acid with sodium. Specific examples include sodium laurylbenzenesulfonate [CAS No.: 25155-30-0, manufactured by FUJIFILM Wako Co., Ltd.], β-naphthalenesulfonic acid formalin condensate sodium salt [CAS No.: 9084-06-4, product name: Demol (registered trademark) RN, manufactured by Kao Corporation], and sodium dioctylsulfosuccinate [CAS No.: 577-11-7, product name: Neocol (registered trademark) SW, manufactured by Daiichi Industry Co., Ltd.].

The cationic or anionic surfactant used in the composition of the present invention is preferably cocamide acetate, lauryltrimethylammonium chloride, sodium cholate, sodium lauryl sulfate, sodium lauryl methylalanine, or sodium β-naphthalenesulfonate formalin condensate. In particular, for plastic containers, lauryltrimethylammonium chloride, sodium cholate, sodium lauryl sulfate, or sodium lauryl methylalanine are more preferably used. For glass containers, lauryltrimethylammonium chloride, cocamide acetate, or sodium β-naphthalenesulfonate formalin condensate are more preferably used.

(Concentration of surfactant)

In the method for improving the storage stability of TARC of the present invention, the concentration of the cationic surfactant or the anionic surfactant is preferably 0.00001 mass% to 1 mass% relative to the composition, more preferably 0.0001 mass% to 1 mass%, further preferably 0.001 mass% to 0.5 mass%, and most preferably 0.001 mass% to 0.1 mass%, based on the stability of the TARC, but is not limited to these.

(TARC adsorption inhibitor)

The TARC adsorption inhibitor of the present invention comprises a cationic surfactant or an anionic surfactant. The TARC adsorption inhibitor comprising a cationic surfactant or anionic surfactant can be added before the TARC solution comes into contact with the storage container. Alternatively, the cationic surfactant or anionic surfactant can be added to the TARC solution in advance as an adsorption inhibitor and then added to the storage container.

Next, the present invention is described in detail with reference to examples. However, these examples do not limit the scope of the present invention. Furthermore, unless otherwise specified, % in this specification represents weight percentage. Furthermore, the product names, ingredient names, and distributors of the surfactants used in the examples are as follows.

‧Emanon (registered trademark) 1112 Ingredient: Polyethylene glycol monolaurate, Kao Corporation

‧Brij (registered trademark) 35 Ingredient: Polyoxyethylene lauryl ether, Kishida Chemical Co., Ltd.

Triton (registered trademark) X-100 Ingredient: Polyethylene glycol mono-p-isooctylphenyl ether, Kishida Chemical Co., Ltd.

‧MEGA-9 Ingredient: N-nonanoyl-N-methyl-D-reduced glucosamine, Tongren Chemical Research Institute Co., Ltd.

‧Sodium Cholate Ingredient Name: Sodium Cholate, FUJIFILM Wako Co., Ltd.

‧SDS Ingredient: Sodium lauryl sulfate, Sigma Aldrich Co., Ltd.

Enagicol (registered trademark) L-30AN Ingredient: Sodium lauryl methylalanine, Lion Specialty Chemicals Co., Ltd.

Demol (registered trademark) RN Ingredient: β-naphthalenesulfonic acid formalin condensate sodium salt, Kao Corporation

Acetamin (registered trademark) 24 Ingredient: Cocoamine Acetate, Kao Corporation

Quartamin (registered trademark) 24P Ingredient: Lauryltrimethylammonium chloride, Kao Corporation

‧CHAPS Ingredient Name: 3-[(3-Cholamidopropyl)dimethylammonio]propanesulfonate, Tongren Chemical Research Institute Co., Ltd.

[Example]

Example 1: Storage Stability Test Results in Plastic Eye Drop Bottles

The storage stability of TARC in plastic eye dropper bottles was tested. The test method and evaluation method are as follows. The following solution was used as the storage solution.

‧PBS (pH 7.2)

‧1% BSA by mass

‧Surfactant (Concentration, product name, and type are listed in Table 1)

(1) Storage conditions

0.5 mL of TARC preservative solution of various concentrations was dispensed into plastic eye dropper bottles and stored at 37°C for 28 days or at 4°C for 28 days. TARC concentrations of 500 pg/mL, 2000 pg/mL, and 10,000 pg/mL were tested.

(2) Measurement method

The assay was performed using a latex immunoassay using two antibodies. The reagent composition and assay method are shown below. The assay was performed using the first and second reagents using a Hitachi automated analyzer.

‧First test agent

100mM MOPS-NaOH (pH 7.5)

500mM NaCl

0.5% BSA

‧Second test reagent

Anti-human TARC monoclonal antibody sensitized latex (2 types)

5mM MOPS-NaOH (pH 7.0)

Furthermore, anti-human TARC monoclonal antibodies were obtained using commercially available TARC antigens using methods known in the industry. Commercially available TARC antigens include: CCL17, thymus and activation regulated chemokine (Shenandoah Biotechnology, Inc.), CCL17/TARC, Human (LifeSpan Bioscience, Inc.), and Human TARC (CCL17) (Abeomics, Inc.). Furthermore, a combination of monoclonal antibodies capable of sandwich assays against TARC antigens was selected using methods known in the industry. The anti-human TARC monoclonal antibody-sensitized latex was prepared using the method described in Japanese Patent Application Laid-Open No. 2017-181377.

First, 120 μL of the first reagent was added to 2.4 μL of TARC preservative solution at each concentration. After warming at 37°C for 5 minutes, 40 μL of the second reagent was added and stirred. The absorbance change over 5 minutes was measured at a primary wavelength of 570 nm and a secondary wavelength of 800 nm. The measured absorbance change was converted to TARC concentration using a calibration curve obtained by measuring a standard substance of known concentration.

(3) Calculation of TARC residual rate (%)

The TARC residual rate (%) was calculated using the following formula based on the TARC concentration in each TARC storage solution after storage at 37°C for 28 days or at 4°C for 28 days.

TARC residual rate (%) = TARC concentration (pg/mL) in each TARC liquid storage solution after storage at 37°C or 4°C for 28 days in the eye drop bottle / TARC concentration (pg/mL) in the newly prepared TARC liquid storage solution × 100

(4) The surfactants and concentrations used under various conditions are shown in Table 1, and the evaluation results are shown in Tables 2 and 3. In addition, the TARC residual rate (%) is calculated based on the average value of three experiments.

[Table 2]

At a TARC concentration of 500 pg/mL, under condition 1, without the addition of a surfactant, the residual rate after 28 days was a very low 47.2%. Under conditions 2-5, which added non-ionic surfactants of the fatty acid ester, alcohol, alkylphenol, or glycosylamide type, and condition 6, which added an amphoteric surfactant, the residual rates after 28 days were 67.7-78.8%, all below 80.0%. In particular, condition 6 contains the compound described in patent document 2, which has been shown to improve the storage stability of insulin. However, the improvement in the storage stability of TARC was minimal. On the other hand, under conditions 7-10, when anionic or cationic surfactants, as discovered in this case, were added, the residual rate after 28 days was over 80% for all surfactants. This is believed to be because under conditions 7-10, the addition of anionic or cationic surfactants prevented TARC from adsorbing to the container walls, or inhibited structural changes in TARC during storage.

[Table 3]

At a TARC concentration of 500 pg/mL, under condition 1 (no surfactant added), the residual rate after 28 days was a relatively low 65.7%. Under conditions 7-10 (adding the anionic or cationic surfactants discovered in this invention), the residual rate after 28 days was 92.0-100.5%, significantly improving the residual rate after 28 days in all conditions. This is believed to be because the addition of the anionic or cationic surfactants under conditions 7-10 prevents TARC from adsorbing to the container walls or inhibits structural changes in TARC during storage.

From the above, it can be seen that the coexistence of anionic or cationic surfactants with TARC improves the storage stability of TARC when stored in plastic eye drop bottles.

Example 2: Storage stability test results in glass vials

The storage stability of TARC in glass vials was tested. The test method and evaluation method are as follows. The following solution was used as the storage solution.

‧PBS (pH 7.2)

‧1% BSA by mass

‧Surfactant (Concentration, product name, and type are listed in Table 4)

(1) Storage conditions

0.5 mL of TARC preservative solution at each concentration was dispensed into glass containers and stored at 10°C for 6 hours. As a control, samples prepared before dispensing into glass containers were also used for the assay. TARC concentrations of 2,000 pg/mL and 10,000 pg/mL were tested.

(2) Measurement method

The measurement was performed using the same method as in Example 1.

(3) Calculation of TARC residual rate (%)

The TARC residual rate (%) was calculated using the following formula based on the TARC concentration in each TARC liquid storage solution after storage at 10°C for 6 hours.

TARC residual rate (%) = TARC concentration (pg/mL) of each TARC liquid storage solution after storage at 10°C in a glass container for 6 hours / TARC concentration (pg/mL) of the TARC liquid storage solution before being dispensed into the glass container × 100

(4) The surfactants and concentrations used under various conditions are shown in Table 4, and the evaluation results are shown in Table 5. In addition, the TARC residual rate (%) is calculated based on the average value of three experiments.

At a TARC concentration of 2000 pg/mL, the residual rate after 6 hours in condition 1, without the addition of a surfactant, was 42.7%, a relatively low result. In conditions 2 to 6, with the addition of nonionic or amphoteric surfactants, the residual rates after 6 hours ranged from 48.6% to 52.5%, showing little improvement.

Under conditions 10-12, when a cationic or anionic surfactant was added, the residual rate after 6 hours improved in all cases, with a particularly significant improvement under condition 12, reaching 93.7%. Similarly, at a TARC concentration of 10,000 pg/mL, the residual rate under conditions 10-12, when a cationic or anionic surfactant was added, improved similarly to that at a TARC concentration of 2,000 pg/mL. In particular, the residual rates under conditions 11 and 12 showed significant improvements, reaching 93.7% and 96.6%, respectively.

The above results indicate that the coexistence of a cationic or anionic surfactant with TARC improves the storage stability of TARC when stored in a glass vial. This is believed to be because, under conditions 10-12, the addition of a cationic or anionic surfactant prevents TARC from adsorbing to the container walls, or inhibits structural changes in TARC during storage.

[Industrial Availability]

According to the present invention, a TARC-containing composition with high storage stability, particularly a TARC-containing calibration sample, can be provided.

Claims (17)

A composition containing TARC (thymus and activation-regulated chemokine), comprising TARC and at least one surfactant selected from the group consisting of cationic surfactants and anionic surfactants, and being in liquid form; wherein the concentration of the TARC relative to the composition is 10 pg/mL to 1 μg/mL; the concentration of the surfactant relative to the composition is 0.00001 mass% to 1 mass%; and the cationic surfactant is an alkylamine salt type or a quaternary ammonium salt type cationic surfactant. The anionic surfactant is a cholic acid type, a sulfate type, a carboxylic acid type, or a sulfonic acid type anionic surfactant. The composition containing TARC as claimed in claim 1 is a calibration sample solution for the determination of TARC. The composition containing TARC as claimed in claim 1 or 2, which is filled in a storage container. The TARC-containing composition of claim 3, wherein the storage container is plastic or glass. The composition containing TARC of claim 1 or 2, wherein the concentration of TARC relative to the composition is 100 pg/mL to 50 ng/mL. In the TARC-containing composition of claim 1 or 2, the concentration of the surfactant is 0.001 mass % to 0.1 mass % relative to the composition. In the composition containing TARC of claim 1 or 2, when the TARC concentration relative to the composition is 500 pg/mL, the residual rate of TARC after storage at 37°C in a plastic container for 28 days is 80% or greater. For the TARC-containing composition of claim 1 or 2, when the TARC concentration relative to the composition is 500 pg/mL, the residual rate of TARC after storage at 4°C in a plastic container for 28 days is 90% or greater. In the composition containing TARC of claim 1 or 2, when the concentration of TARC in the composition is 2000 pg/mL, the residual rate of TARC after storage at 10°C in a glass container for 6 hours is 55% or more. The TARC-containing composition of claim 1 or 2, wherein the cationic surfactant is lauryltrimethylammonium chloride or cocamine acetate, and the anionic surfactant is sodium cholate, sodium lauryl sulfate, sodium lauryl methylalanine, or sodium β-naphthalenesulfonate formalin condensate. A method for determining TARC, comprising using the composition of any one of claims 1 to 10. A TARC assay kit comprising the composition of any one of claims 1 to 10. A method for improving the storage stability of TARC comprises contacting the TARC with a solution containing at least one surfactant selected from the group consisting of cationic surfactants and anionic surfactants, and adjusting the concentration of the TARC relative to the solution to be between 10 pg/mL and 1 μg/mL. The concentration of the surfactant relative to the solution is between 0.00001 mass% and 1 mass%. The cationic surfactant is an alkylamine salt-type or quaternary ammonium salt-type cationic surfactant, and the anionic surfactant is a cholic acid-type, sulfate-type, carboxylic acid-type, or sulfonic acid-type anionic surfactant. The method for improving the storage stability of TARC according to claim 13, comprising the step of adjusting the concentration of the TARC in the solution to be between 100 pg/mL and 50 ng/mL. The method for improving the storage stability of TARC according to claim 13 or 14, wherein the concentration of the surfactant is 0.001 mass % to 0.1 mass % relative to the solution. The method for improving the storage stability of TARC according to claim 13 or 14, wherein the cationic surfactant is lauryltrimethylammonium chloride or cocamide acetate, and the anionic surfactant is sodium cholate, sodium lauryl sulfate, sodium lauryl methylalanine, or sodium β-naphthalenesulfonate formalin condensate. A method for preventing TARC from being adsorbed onto a container in a solution containing TARC, the method comprising adding a TARC adsorption inhibitor, the TARC adsorption inhibitor containing as an active ingredient at least one surfactant selected from the group consisting of cationic surfactants and anionic surfactants, the concentration of TARC being 10 pg/mL to 1 μg/mL relative to the solution, the concentration of the surfactant being 0.00001 mass% to 1 mass% relative to the solution, and the cationic surfactant being an alkylamine salt type or a quaternary ammonium salt type cationic surfactant. The anionic surfactant is a cholic acid type, a sulfate type, a carboxylic acid type, or a sulfonic acid type anionic surfactant.