Classifications

• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
  • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
  • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
  • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
  • G01N33/6863—Cytokines, i.e. immune system proteins modifying a biological response such as cell growth proliferation or differentiation, e.g. TNF, CNF, GM-CSF, lymphotoxin, MIF or their receptors
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
  • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
  • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
  • G01N33/96—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood or serum control standard
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
  • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
  • G01N2333/52—Assays involving cytokines
  • G01N2333/521—Chemokines

Definitions

• the present invention relates to a composition containing TARC and a diluent for the composition containing TARC.
• the present invention also relates to a method for suppressing carryover of TARC and an antiadsorption agent for TARC.
• the present invention also relates to methods for the continuous analysis of two or more samples containing TARC.
• Thymus and activation-regulated chemokine (hereinafter sometimes referred to as TARC) is C—C chemokine ligand 17 (CCL17) and is a kind of chemokine with leukocyte migration activity.
• TARC attracts Th2 cells, which are one of lymphocytes, to the lesion site, and causes IgE production and eosinophil infiltration/activation. It is believed that TARC exacerbates the symptoms of atopic dermatitis by enhancing allergic reactions in this way (Non-Patent Document 1).
• TARC matches well with the severity of atopic dermatitis, It is believed that it more sensitively reflects disease activity (Non-Patent Document 2). Therefore, by using TARC as a biomarker, it is possible to objectively and quickly grasp the severity of atopic dermatitis and determine its effect when selecting or changing a therapeutic agent for atopic dermatitis.
• a calibration sample is a sample containing a component to be measured, which is used for applications such as an internal standard or concentration calibration standard (calibrator).
• the calibration sample is preferably in the form of a fluid solution (hereinafter sometimes referred to as "liquid") for ease of operation.
• the desired items for the calibration sample include biological activity (for example, antigenicity to specific antibodies, binding activity to specific binding partners possessed by antibodies and lectins, physiological activity possessed by peptide hormones, enzymatic activity, etc.). , three-dimensional structure as a protein for supporting each of the above activities), prevention of adsorption to containers, maintenance of antiseptic ability, and the like.
• Methods for preserving a calibration sample intended for use in an immunoassay method in liquid form include a method of stabilizing an antigen by allowing casein and/or whey protein to coexist in a calibration sample (Patent Document 1); A method of stabilizing insulin in the presence of an acid amide derivative (Patent Document 2) and a method of stabilizing soluble interleukin-2 receptor (sIL-2R) in the presence of a chelating agent (Patent Document 3) are known. ing.
• Patent Document 4 a cationic surfactant or an anionic surfactant
• Patent Document 5 a sugar fatty acid ester type nonionic surfactant
• Patent Document 6 2-methacryloyloxyethyl A method using a polymer having phosphorylcholine as a structural unit
• the sample When measuring components in a sample using an automatic analyzer, the sample is dispensed into a dilution cell or a reaction cell by a sample probe.
• the sample probe is a fine tube for sucking up a required amount of a sample (calibration sample, sample, sample diluted with a diluent) and putting it into a reaction cell.
• the sample probe operates based on a program unique to each automatic analyzer. At the time of concentration calibration, sampling is performed from the low concentration side, so even though the sample probe is rinsed with water for each sample, many devices do not have a program for washing with detergent. In addition, some devices do not even rinse the sample probe with water after each sample, but are programmed to rinse or clean with detergent only at the end of sampling of all calibration standards.
• a high-concentration reference material for calibration should be used.
• Zero-concentration or low-concentration calibration samples are sampled without washing the aspirated sample probe. Antigens contained in high-concentration calibration samples tend to adhere to the sample probe, and if they cannot be rinsed off with water, contamination of the next sample, ie, carryover, occurs. As described above, there is a problem that the occurrence of carryover at the time of concentration calibration makes it impossible to create an accurate calibration curve, making it impossible to obtain an accurate measurement value of the sample.
• the sample probe when measuring samples, the sample probe is usually rinsed with water for each sample, so carryover can occur in the same way as with calibration samples. Carryover can be avoided by adding (carryover avoidance setting).
• some automatic analyzers are not equipped with the above-mentioned cleaning mechanism, and there is a demerit that the processing capacity is lowered due to the cleaning of the sample probe.
• the present inventors have discovered that when TARC is continuously quantified, the TARC antigen is adsorbed to the sample probe and carryover occurs in subsequent samples to be quantified.
• the present inventors diligently studied a method that does not cause carryover.
• the inventors have found that the occurrence of carryover can be prevented by adopting the following configuration, and have completed the present invention.
• the present invention is as follows. ⁇ 1> A composition comprising TARC (Thymus and activation-regulated chemokine), (A) TARC, and (B) one or more components selected from the group consisting of acidic amino acids, basic amino acids, acidic polymers having an average molecular weight of 4,000 to 1,200,000, and salts thereof; and the composition, which is liquid.
• composition according to ⁇ 1> which is a calibration sample solution for measuring TARC.
• C The composition according to ⁇ 1> or ⁇ 2>, further comprising a buffer.
• ⁇ 4> The composition according to any one of ⁇ 1> to ⁇ 3>, wherein the acidic polymer and its salt are sodium polystyrenesulfonate, naphthalenesulfonic acid condensate, sodium polyacrylate, or polyacrylic acid.
• concentration of component (B) is 0.0001 to 20% by mass relative to the composition.
• ⁇ 6> (B) dilution of a TARC-containing sample containing one or more components selected from the group consisting of acidic amino acids, basic amino acids, acidic polymers having an average molecular weight of 4,000 to 1,200,000, and salts thereof; liquid.
• the diluent according to ⁇ 6>, wherein the acidic polymer and its salt are sodium polystyrenesulfonate, naphthalenesulfonic acid condensate, sodium polyacrylate, or polyacrylic acid.
• C The composition according to ⁇ 6> or ⁇ 7>, further comprising a buffer.
• ⁇ 9> The diluent according to any one of ⁇ 6> to ⁇ 8>, wherein the concentration of the component (B) is 0.0001 to 20% by mass relative to the diluent.
• a method for suppressing carryover of TARC comprising: ⁇ 11> The method for suppressing carryover of TARC according to ⁇ 10>, wherein the acidic polymer and its salt are sodium polystyrenesulfonate, naphthalenesulfonic acid condensate, sodium polyacrylate, or polyacrylic acid.
• ⁇ 12> The method for suppressing carryover of TARC according to ⁇ 10> or ⁇ 11>, wherein the solution is a buffer solution.
• ⁇ 13> The method for suppressing TARC carryover according to any one of ⁇ 10> to ⁇ 12>, wherein the carryover is the carryover in the sample probe.
• ⁇ 14> The method for suppressing carryover of TARC according to any one of ⁇ 10> to ⁇ 13>, wherein the concentration of the component (B) is 0.0001 to 20% by mass with respect to the solution.
• a method for the continuous analysis of two or more samples containing TARC comprising: (a) aspirating a first sample containing TARC with a sample probe; (b) expelling said first sample from the sample probe; (c) measuring signal intensity in said first sample; and (d) rinsing the walls of the sample probe with a rinsing solution; (e) aspirating a second sample containing TARC with the sample probe; and (f) measuring signal intensity in the second sample;
• the first sample and the second sample are (B) one or more selected from the group consisting of acidic amino acids, basic amino acids, acidic polymers having an average molecular weight of 4,000 to 1,200,000, and salts thereof 0.0001% by mass to 20% by mass of each component with respect to the first sample and the second sample, and The method of continuous analysis, wherein the concentration of TARC in the first sample containing TARC is higher than the concentration of TARC in the second sample containing TARC.
• the present invention it is possible to provide a calibration sample and specimen diluent in which carryover of the TARC antigen is less likely to occur. Therefore, according to the present invention, it is possible to accurately quantify TARC in a biological sample, and accurately grasp the severity of atopic dermatitis.
• TARC means Thymus and activation-regulated chemokine (CCL17).
• TARC is a kind of chemokine with leukocyte migration activity.
• TARC has the function of attracting Th2 cells, which are one of lymphocytes, to the lesion site and infiltrating and activating IgE production or eosinophils.
• Th2 cells which are one of lymphocytes
• IgE production or eosinophils By using TARC as a biomarker, the severity can be objectively and quickly grasped when selecting or changing therapeutic agents for atopic dermatitis.
• TARC can be performed using known techniques, such as immunological techniques.
• Immunological techniques include ELISA, enzyme immunoassay, surface plasmon resonance, latex agglutination immunoassay (LTIA), chemiluminescence immunoassay, electrochemiluminescence immunoassay, fluorescent antibody method, radioimmunoassay, Western blotting, immunochromatography, high performance liquid chromatography (HPLC method) and the like can be mentioned.
• TARC contained in the composition of the present invention a commercially available one may be used, or one produced or purified by oneself may be used.
• TARC contained in the composition of the present invention one produced in vitro or one extracted from a living body may be used.
• the concentration of TARC contained in the composition of the present invention is not limited to the following, but is preferably 10 pg/mL to 1 ⁇ g/mL, more preferably 50 pg/mL to 500 ng/mL, relative to the composition. , more preferably 100 pg/mL to 100 ng/mL, most preferably 100 pg/mL to 50 ng/mL.
• composition of the present invention comprises one or more components (hereinafter simply component (sometimes referred to as (B)). These components ionize in aqueous solution and exhibit a negative or positive charge. Being ionized in an aqueous solution and exhibiting a negative charge means having more acidic functional groups such as a sulfo group than basic functional groups such as an amino group, and exhibiting a negative charge when dissolved in water. Being ionized in an aqueous solution and exhibiting a positive charge means having more basic functional groups than acidic functional groups and exhibiting a positive charge when dissolved in water.
• the acidic functional group includes a sulfo group, a phospho group, a carboxyl group, and the like.
• a sulfo group or a carboxyl group is preferred.
• Basic functional groups include primary amino groups, secondary amino groups, tertiary amino groups, amino groups such as quaternary ammonium salts, amide groups, pyridyl groups, pyridine groups, pyrrolidone groups, imidazole groups, Or an imine group etc. are mentioned. Amino groups are preferred.
• Acidic amino acids, acidic polymers having a weight average molecular weight of 4,000 to 1,200,000, or salts thereof include L-glutamic acid, L-aspartic acid, sodium polystyrene sulfonate (CAS registration number 25704-18-1), Sodium polyacrylate (CAS Registry Number 9003-04-7), naphthalenesulfonic acid condensate (CAS Registry Number 9084-06-4), or polyacrylic acid (CAS Registry Number 9003-01-4) can be used.
• naphthalenesulfonic acid condensate one manufactured and sold by Kao Corporation as Demol (registered trademark) RN can be used.
• the amino acids referred to in this specification may be ⁇ -amino acids that are structural units of living organisms, or may be non-natural amino acids.
• amino acids include L-forms, D-forms, and mixtures thereof.
• Basic amino acid or its salt As basic amino acids or salts thereof, L-lysine, L-arginine, or salts thereof can be used.
• Salts of acidic amino acids, salts of basic amino acids, and salts of acidic polymers having a weight average molecular weight of 4,000 to 1,200,000 are not limited as long as the effects of the present invention can be obtained.
• Acidic amino acids, basic amino acids, or acidic polymers having a weight average molecular weight of 4,000 to 1,200,000 include L-glutamic acid, L-aspartic acid, sodium polystyrenesulfonate, naphthalenesulfonic acid condensates, and polyacrylic acid. is preferred, and L-glutamic acid, L-aspartic acid, sodium polystyrene sulfonate, or naphthalenesulfonic acid condensate (Demol RN) is more preferred.
• As the acidic polymer those having an average molecular weight (Average MW) of, for example, 4,000 to 1,200,000, preferably 5,000 to 1,000,000 can be used.
• the weight average molecular weight (MW) is preferably 70,000 to 1,000,000.
• a weight average molecular weight (MW) can be measured using size exclusion chromatography (SEC:Size Exclusion Chromatography).
• SEC:Size Exclusion Chromatography size exclusion chromatography
• any polymer obtained by polymerizing a monomer having an acidic group can be used without limit.
• the acidic polymer is preferably an acidic polymer obtained by polymerizing a monomer having a carboxyl group or a sulfo group.
• the concentration of component (B) is, for example, 0.0001 to 20% by weight, preferably 0.001 to 10% by weight, based on the composition.
• the concentration is more preferably 0.05 to 10% by mass, more preferably 0.1 to 5% by mass, most preferably 0.5 to 1% by mass, relative to the composition. %.
• the concentration is more preferably 0.001 to 1% by mass, more preferably 0.001 to 0.5% by mass, most preferably 0.002 to It is 0.1% by mass.
• the concentration is, for example, 1 to 20% by mass, preferably 3 to 18% by mass, more preferably 5 to 15% by mass, most preferably 5 to 15% by mass, relative to the composition. 10% by mass.
• component (B) and TARC is not particularly limited as long as the effects of the present invention can be obtained.
• buffer means a solution that has a pH buffering effect.
• the buffers used in the present invention are not limited to the following, but examples include PBS (phosphate buffered saline), MES (2-(N-morpholino) ethanesulfonic acid), PIPES (piperazine-N,N'- bis(2-ethanesulfonic acid), ACES (N-(2-Acetamido)-2-aminoethanesulfonic acid), ADA (N-(2-Acetamido)iminodiacetic acid), Bis-Tris (2,2-Bis(hydroxyethyl)- (iminotris)-(hydroxymethyl)-methane), Tris (tris(hydroxymethyl)aminomethane), MOPS (3-Morpholinopropanesulfonic acid), HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), citric acid buffer,
• the buffer used in the present invention is preferably PBS.
• the concentration of the buffer solution is not particularly limited as long as the effect of the present invention can be obtained, but is, for example, 1-500 mM, 5-400 mM, 10-300 mM, or 50-200 mM.
• the pH of the buffer used in the present invention is, for example, 4.0-11.0, 5.0-10.0, 6.0-9.0, 6.5-8.0, or 7.0. ⁇ 8.0.
• the pH can be adjusted using pH adjusting reagents well known to those skilled in the art, such as sodium hydroxide or hydrochloric acid.
• the content of the buffer is, for example, 90% by mass or more, 92% by mass or more, 95% by mass or more, 97% by mass or more, or 99% by mass or more relative to the composition. can.
• compositions of the present invention are preferably packaged in storage containers.
• the material of the storage container is not particularly limited as long as the effects of the present invention can be obtained and can be sealed, but at least part or all of the contact portion with the composition is plastic [e.g., olefin resin, styrene resin , acrylic resin, polyester resin, polycarbonate resin, fluorine resin, chlorine resin (polyvinyl chloride, etc.), polyamide resin, polyacetal resin, polyphenylene ether resin (modified polyphenylene ether, etc.), polyarylate, polystyrene sulfone , polyimide-based resins, cellulose-based resins (cellulose acetate, etc.), hydrocarbon-based resins (including halogen atom-substituted products), etc.], metals (aluminum, etc.), glass, and the like.
• plastic or glass is preferable from the viewpoint of production, transportation, and storage of the calibration sample.
• plastics e.g.,
• the storage container may be composed of a single material or two or more materials, but is preferably composed of a single material.
• the storage container can include a container body portion and a cap.
• the container body portion and the cap may be made of different materials.
• the storage container, particularly the body portion preferably has transparency to the extent that the liquid content can be seen from the outside.
• the form of the storage container may be either hard type or soft type, and examples thereof include ampoules, vials, soft bags, injection-type containers, and glass bottles. From the viewpoint of ease of use and stability of TARC, the storage container is preferably in the form of a plastic eyedropper bottle, particularly in the form of a cylindrical eyedropper bottle, including a container body portion and a cap portion.
• composition The composition of the present invention can be used as a calibration sample solution in the measurement of TARC.
• the calibration sample solution means a sample solution containing the substance to be measured at a certain concentration, which is used to accurately measure the substance to be measured. This includes internal standard substances.
• the supply form of the composition of the present invention includes a form in which the TARC and the component (B) are mixed in a solvent and brought into a solution state in advance.
• "using a composition” means using a composition to accurately measure TARC. substances, calibrators, controls, internal standards, etc.).
• the pH of the composition of the present invention is, for example, 4.0-11.0, 5.0-10.0, 6.0-9.0, 6.5-8.0, or 7.0-8. is 0.
• the pH can be adjusted using pH adjusting reagents well known to those skilled in the art, such as sodium hydroxide or hydrochloric acid.
• composition of the composition of the present invention is not particularly limited as long as it does not impair the effects of the present invention. If TARC is measured by an immunoassay method, it should not impair the effects of the present invention and interfere with all or part of the reaction that constitutes the assay system, such as antigen-antibody reaction or enzymatic reaction.
• components commonly used in immunoassay methods such as components that promote antigen-antibody reactions (polymers such as polyethylene glycol and polyvinylpyrrolidone, etc.), glycoproteins and peptides (BSA, casein, etc.), amino acids, salts ( Sodium chloride, potassium chloride, etc.), saccharides (sucrose, cyclodextrin, etc.), preservatives (sodium azide, ProClin300, etc.), other anti-adsorption agents (Lipidure, etc.), etc. are appropriately selected and used according to the purpose. be able to.
• polymers such as polyethylene glycol and polyvinylpyrrolidone, etc.
• BSA glycoproteins and peptides
• amino acids Sodium chloride, potassium chloride, etc.
• saccharides saccharides
• preservatives sodium azide, ProClin300, etc.
• other anti-adsorption agents Lipidure, etc.
• the term "carryover" means that when the first sample is fractionated, the TARC antigen in the sample remains adsorbed to the wall surface of the sample probe, etc., and the reaction after the second sample that is sampled next. This means that the cells are flooded with the TARC antigen from the first sample.
• carryover occurs, the signal intensity of the second and subsequent samples becomes higher than the signal intensity originally obtained, making it impossible to obtain an accurate quantification value.
• the inventors have found that carryover occurs even when the TARC is in contact with the sample probe only for a very short time, as described above. Use of the compositions of the present invention can prevent carryover and improve the accuracy of TARC measurements.
• a sample probe means a fine tube for sucking up a required amount of specimen from a container such as a blood collection tube and putting it into a reaction cell.
• Sample probes include both those for automated measurement equipment and those for manual use.
• the sample probe is preferably made of metal, more preferably stainless steel.
• the sample for TARC measurement is not particularly limited as long as TARC can be measured, but blood, serum, or plasma is preferably used.
• the sample may be appropriately pretreated as necessary.
• the sample is preferably a biological sample taken from humans.
• TARC reagent kit The composition of the invention can be provided in the form of a TARC reagent kit containing the composition of the invention and other reagents.
• reagent kits include reagent kits using immunological techniques. Immunological techniques include ELISA, enzyme immunoassay, surface plasmon resonance, latex agglutination immunoassay (LTIA), chemiluminescence immunoassay, electrochemiluminescence immunoassay, fluorescent antibody method, radioimmunoassay, Western blotting, immunochromatography, high performance liquid chromatography (HPLC method) and the like can be mentioned.
• the reagent kit can be used for selecting a treatment method or drug for atopic dermatitis, and for grasping the severity of atopic dermatitis when determining the effect of treatment.
• the reagent kit can also include instructions for use and the like.
• the reagent kit may contain optional components such as stabilizers, pH adjusters, reaction vessels, and the like.
• sample containing TARC sample containing TARC
• a diluent of the invention can be used to dilute a sample containing TARC.
• Samples containing TARC include samples for measuring TARC (blood, serum, plasma, etc.), standards, calibrators, controls, and internal standards.
• the composition other than not containing TARC is the same as the above "1.
• Composition containing TARC is the same as the above "1.
• the diluent of the present invention preferably has a concentration of TARC after dilution of 10 pg/mL to 1 ⁇ g/mL, more preferably 50 pg/mL to 500 ng/mL, still more preferably 100 pg/mL to 100 ng/mL, most preferably 100 pg/mL to 50 ng/mL can be used.
• the method for suppressing carryover of TARC of the present invention includes contacting TARC and component (B) in a solution.
• TARC may be added after component (B) is added to the solution, or component (B) may be added after TARC is added to the solution.
• the solution is a buffer solution, more preferably PBS.
• TARC as component (A) and the compound found in the present application as component (B) may be brought into contact with each other in a sample probe. After contacting with, the prepared TARC-containing composition may be transferred to the sample probe.
• a method of suppressing carryover of TARC of the present invention can comprise retaining a prepared composition comprising TARC in a sample probe.
• "Holding the prepared composition containing TARC in the sample probe” means that the composition containing TARC is in contact with the wall surface of the sample probe, and other operations, such as dispensing a sample Movement of the probe and the like may be performed in parallel.
• the retention time of TARC in the sample probe is not limited as long as the effects of the present invention can be obtained.
• the upper limit is 1 hour or less, 30 minutes or less, 15 minutes or less, 10 minutes or less, 5 minutes or less, 1 minute or less, 30 seconds or less, 10 seconds or less, 5 seconds or less, 3 seconds or less, or 2 seconds or less. can be done.
• Specific retention time ranges include 0.1 seconds to 1 hour, 0.1 seconds to 30 minutes, 0.1 seconds to 15 minutes, 0.1 seconds to 10 minutes, 0.1 seconds to 1 minute, It can be 0.2 seconds to 30 seconds, 0.2 seconds to 10 seconds, 0.5 seconds to 5 seconds, or 0.5 seconds to 3 seconds.
• the TARC carryover suppression method of the present invention can optionally further comprise one or more of the following steps.
• the first sample containing TARC and the second and subsequent samples containing TARC both contain component (B).
• the component (B) contained in the first sample and the component (B) contained in the second sample are preferably the same.
• Component (B) may be added to the first sample and the second sample as a sample diluent prior to measurement, or may contain component (B) from the beginning.
• the first and second samples may be either samples for measuring TARC (blood, serum, plasma, etc.) or calibration samples (standard substances, calibrators, controls, etc.).
• the first and second samples are preferably calibration samples (standard substances, calibrators, controls, etc.) for measuring TARC.
• “Aspirating with the sample probe" in steps (a) and (e) means that the person performing the analysis manually aspirates the first sample, and the person performing the analysis turns on the switch so that the automatic analyzer and automatically aspirating the first sample.
• steps other than (a) As an automatic analyzer, latex turbidimetric immunoassay (LTIA method), electrochemiluminescence immunoassay (ECL method), enzyme-linked immunosorbent assay (ELISA), chemiluminescence immunoassay, immunofluorescence assay, etc.
• An automatic analyzer or the like can be used.
• steps (a) to (g) steps (a), (b), (d), (e), and (f) are performed in the stated order.
• the timing of performing (c) and (g) can be freely set according to the program of the automatic analyzer to be used.
• the substance that generates the signal can be appropriately selected according to the type of analysis method.
• Signal-generating substances include, for example, metal complexes, enzymes, insoluble particles, fluorescent substances, chemiluminescent substances, biotin, avidin, radioactive isotopes, colloidal gold particles, latex particles or colored latex.
• the signal is not limited as long as the concentration of TARC in the sample can be measured based on the intensity of the signal, and fluorescence, absorbance, scattered light intensity, or the like can be appropriately selected according to the type of analysis method.
• the TARC antigen is attached to the wall surface of the sample probe after the first sample is ejected. Therefore, if there is no washing step with a washing agent or the like after discharging the first sample, carryover of the TARC antigen will occur in the second and subsequent samples to be measured next.
• rinsesing the wall surface of the sample probe means that the sample probe is rinsed by the frictional force, shear stress, etc. generated when the rinsing liquid flows without using the action of rubbing or polishing with a brush or the like. means to wash away the walls of
• the rinse often consists essentially of water, and more preferably the rinse is essentially free of cleaning agents. This is because if the rinsing solution contains a detergent, the detergent remaining in the sample probe must be further rinsed with water, extending the time required for the rinsing process.
• the rinsing liquid contains a detergent
• the content of the detergent in the rinsing liquid is preferably 0.1% by mass or less, more preferably 0.01% by mass or less, It is more preferably 0.001% by mass or less, and most preferably contains no detergent.
• detergent is meant a substance that has the ability to release proteins from the walls of the sample probe on the basis of chemical action. Detergents specifically include acids, alkalis, surfactants, enzymes, and the like.
• the concentration of TARC in the first sample containing TARC is higher than the concentration of TARC in the second and subsequent samples containing TARC.
• the concentration of TARC in the first sample containing TARC is higher than the concentration of TARC in the second sample containing the TARC, the signal intensity in the second and subsequent samples is originally reduced due to the influence of carryover of the TARC antigen. It may become high compared with the obtained signal intensity, and it may not be possible to obtain an accurate quantification value.
• the concentration of TARC in the first sample is, for example, 500 pg/mL or higher, 1,000 pg/mL or higher, 5,000 pg/mL or higher, 10,000 pg/mL or higher, 15,000 pg/mL or higher, immediately before being discharged from the sample probe. mL or greater, or 20,000 pg/mL or greater.
• the concentration of TARC in the second sample may be lower than that of the first sample immediately before it is discharged from the sample probe.
• mL 0 pg/mL to 1000 pg/mL, 0 pg/mL to 700 pg/mL, 0 pg/mL to 600 pg/mL, 0 pg/mL to 500 pg/mL, 0 pg/mL to 300 pg/mL, or 0 pg/mL to 100 pg/mL is.
• the above concentration is the concentration after dilution when the specimen diluent is added.
• the inclusion of the component (B) in the first sample can prevent the influence of carryover of the TARC antigen in the second and subsequent samples. Therefore, the concentration of TARC can be measured with high accuracy.
• the TARC antigen carryover suppression method of the present invention can optionally further comprise one or more of the following steps. ⁇ Based on the signal intensities of the first and second and subsequent samples, calculate the concentration of TARC in each of the first and second and subsequent samples. Prepare a calibration curve based on the signal intensity. ⁇ Wash the sample probe with a washing solution containing a detergent at the start and/or at the end of sample analysis. It means when all the samples to be analyzed have been finished.
• the antiadsorption agent for TARC antigen of the present invention contains a compound exhibiting a positive or negative charge as an active ingredient.
• Embodiments of the antiadsorption agents for TARC antigens of the present invention are the same as those described above.
• continuous analysis of two or more samples means continuous analysis of two or more samples by a fully automatic analyzer.
• the mechanism for washing the wall surface of the sample probe with a cleaning agent between each sample analysis is not excluded.
• the continuous analysis method of two or more samples containing the TARC antigen of the present invention can optionally include contacting component (A) TARC and component (B) in solution.
• the step can be, for example, a step of diluting the first sample containing TARC and/or the second and subsequent samples with a specimen diluent containing component (B).
• ⁇ Lipidure (registered trademark) series manufactured by NOF Corporation ⁇ Acetamine (registered trademark) 24
• Ingredient name coconut amine acetate, manufactured by Kao Corporation ⁇ Demoll (registered trademark) RN-L
• Measurement method 1-1 Measurement Reagents An LTIA measurement reagent for TARC measurement, consisting of the first reagent and the second reagent shown below, was prepared. (1) First reagent 100 mM MOPS-NaOH (pH 7.5) 500 mM NaCl 0.5% BSA (2) Second reagent Anti-human TARC monoclonal antibody-sensitized latex (2 types) 5 mM MOPS-NaOH (pH 7.0) The anti-human TARC monoclonal antibody was obtained using a commercially available TARC antigen by a method well known to those skilled in the art.
• TARC antigens included CCL17, thymus and activation regulated chemokine (Shenandoah Biotechnology, Inc.), CCL17/TARC, Human (LifeSpan Bioscience, Inc.), Human TARC (CCL17) (Abeomics, Inc.). Furthermore, a combination of monoclonal antibodies capable of sandwich assay against the TARC antigen was selected by a method well known to those skilled in the art. Anti-human TARC monoclonal antibody-sensitized latex was prepared with reference to the method described in JP-A-2017-181377.
• Sample A TARC antigen was dissolved in a diluent having the following composition to prepare a low-concentration sample (Sample A, 800 pg/mL level) and a high-concentration sample (Sample B, 20,000 pg/mL level). In addition, physiological saline was prepared separately.
• the first reagent and the second reagent were combined, and the TARC concentration in the sample was measured using a Hitachi automatic analyzer 3500 (sample probe made of stainless steel). Specifically, 120 ⁇ L of the first reagent was added to 2.4 ⁇ L of the sample, and the mixture was incubated at 37° C. for 5 minutes, then 40 ⁇ L of the second reagent was added and stirred. Absorbance changes associated with aggregate formation were then measured at a dominant wavelength of 570 nm and a minor wavelength of 800 nm over a period of 5 minutes. Measurement sensitivity was calculated from the amount of change in absorbance.
• Table 1 shows the results when component (B) was not added to the calibrator diluent.
• the measurement sensitivity of the first set of 5 measurements for sample A (800 pg/mL level) was 20.9 to 22.1 mAbs, and the average measurement sensitivity was 21.4 mAbs.
• sample B (20,000 pg/mL level) was measured twice, and then sample A was measured again five times (second set).
• the sensitivity of the first measurement in the second set was 38.3 mAbs, which was much higher than the average sensitivity of the first set, and the relative ratio was poor at 178.5%.
• Tables 2 to 6 show the results when various components were added to the diluted solution so that the final concentrations shown in the table were obtained.
• the table shows an excerpt of the relative ratio of the sensitivity of each measurement of the second set to the average sensitivity of the five measurements of the first set.
• Table 2 shows the results of adding acidic amino acids
• Table 3 shows the results of adding basic or neutral amino acids.
• “A” is a very good result with the relative ratio of the 1st time of the 2nd set within 90 to 110%
• “B” is the relative ratio of the 1st time of the 2nd set is 90 ⁇
• the second relative ratio was within 90-110%, which was a good result.
• sodium polystyrene sulfonate (Example 8: Mw 70,000, Example 9: Mw 200,000, Example 10: Mw 1,000,000), which is a polymer compound obtained by polymerizing a monomer having a sulfo group, and Demol RN (Example 11).
• the relative ratio of the sensitivity of the first measurement of the second set to the average measurement sensitivity of the five measurements of the first set is 99.1 to 104.9%, and the carryover of sample B can be suppressed.
• rice field Sodium polyacrylate (Example 12, molecular weight 22,000-70,000) and polyacrylic acid (Examples 13-15, molecular weight 5,000-100 10,000) also found a certain effect.
• acidic polymers such as sodium polystyrene sulfonate (molecular weight 70,000 to 1,000,000), naphthalenesulfonic acid condensate (Demol RN), sodium polyacrylate (molecular weight 22,000 to 70,000), and polyacrylic acid (molecular weight 5,000 to 1,000,000) was added to the diluent, it was found that the carryover caused by adsorption of the TARC antigen to the sample probe could be suppressed. Therefore, when an acidic polymer is added to the diluent, even if a sample with a low TARC antigen concentration is measured immediately after a sample with a high TARC antigen concentration is measured, the TARC antigen concentration is low. It was considered possible to measure the concentration of the sample with high accuracy.
• DEMO RN Although DEMO RN also has surface activity, it is treated as an acidic polymer in this specification because it has sulfonic acid in its molecule.
• Table 5 shows the study results of compounds with positive charges such as amino groups or quaternary ammonium salts in the molecule.
• the evaluation criteria are the same as those in Tables 2 and 3, but in both cases, no effect of addition was observed with respect to non-addition (Comparative Example 1).

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