[go: up one dir, main page]

WO2023210671A1 - Copolymère, film polymère, dispositif de mesure et support de mesure - Google Patents

Copolymère, film polymère, dispositif de mesure et support de mesure Download PDF

Info

Publication number
WO2023210671A1
WO2023210671A1 PCT/JP2023/016392 JP2023016392W WO2023210671A1 WO 2023210671 A1 WO2023210671 A1 WO 2023210671A1 JP 2023016392 W JP2023016392 W JP 2023016392W WO 2023210671 A1 WO2023210671 A1 WO 2023210671A1
Authority
WO
WIPO (PCT)
Prior art keywords
copolymer
independently
substance
structural unit
present disclosure
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/JP2023/016392
Other languages
English (en)
Japanese (ja)
Inventor
秀治 栗岡
ちさと 小村
剛 安藤
広治 網代
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Kyocera Corp
Original Assignee
Kyocera Corp
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Kyocera Corp filed Critical Kyocera Corp
Priority to JP2024517362A priority Critical patent/JPWO2023210671A1/ja
Publication of WO2023210671A1 publication Critical patent/WO2023210671A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
    • C08F293/00—Macromolecular compounds obtained by polymerisation on to a macromolecule having groups capable of inducing the formation of new polymer chains bound exclusively at one or both ends of the starting macromolecule
    • 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
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/544—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being organic

Definitions

  • the present disclosure relates to a copolymer used to form a polymer membrane used in a measuring device.
  • the present disclosure also relates to a polymer membrane obtained using the copolymer, and a measuring device or a measuring carrier provided with the polymer membrane.
  • a surface acoustic wave sensor is disclosed in Patent Document 1 as an example of a measuring device that measures the concentration of a substance to be measured (for example, a biomolecule).
  • a measuring device includes a detection section on which a substance (for example, an antibody) that interacts with a substance to be measured contained in a specimen is immobilized.
  • a polymer film is often formed in the detection section in order to immobilize a substance that interacts with the substance to be measured contained in the sample.
  • the copolymer according to one embodiment includes two hydrophilic blocks A containing at least one structural unit (a) among the structural units (a) represented by the following formulas (1) to (6), (In the formula, R 1 to R 7 are each independently H or CH 3 , X 1 to X 4 are each independently O or NH, and p 1 to p 5 are each independently 1 to 3 The following integers are shown, and q 1 to q 2 are each independently an integer between 1 and 5.) Between the two hydrophilic blocks A, a hydrophobic block B containing at least one structural unit (b) among the structural units (b) represented by the following formulas (7) and (8), (In the formula, R 8 and R 10 are each independently H or CH 3 , R 9 , R 11 and R 12 are each independently an alkyl group having 1 to 6 carbon atoms, and X 5 and 6 is O or NH, and r is an integer from 1 to 3.)
  • the hydrophobic block B is a copolymer having disulfide bonds between the structural
  • the polymer membrane according to one embodiment includes a hydrophilic block A containing at least one structural unit (a) among structural units (a) represented by the following formulas (1) to (6); (In the formula, R 1 to R 7 are each independently H or CH 3 , X 1 to X 4 are each independently O or NH, and p 1 to p 5 are each independently 1 to 3 The following integers are shown, and q 1 to q 2 are each independently an integer between 1 and 5.) A hydrophobic block B containing at least one structural unit (b) among the structural units (b) represented by the following formulas (7) and (8), (In the formula, R 8 and R 10 are each independently H or CH 3 , R 9 , R 11 and R 12 are each independently an alkyl group having 1 to 6 carbon atoms, and X 5 and 6 is O or NH, and r is an integer from 1 to 3.)
  • the end of the hydrophobic block B opposite to the side bonded to the hydrophilic block A is a polymer membrane containing a cop
  • FIG. 1 is a schematic diagram showing an example of a measuring device according to the present disclosure. It is a top view showing the sensor with which the above-mentioned measuring device is provided.
  • FIG. 2 is a conceptual diagram showing a polymer included in the detection section 23 shown in FIG. 1.
  • FIG. FIG. 2 is a plan view showing an example of a measurement carrier according to the present disclosure. It is a graph showing the relationship between the unit ratio (CBMA1/BMA) in the triblock copolymer and the amount of non-specific adsorption according to the present disclosure. It is a graph showing the relationship between the average degree of polymerization of CBMA1 in the triblock copolymer according to the present disclosure and the amount of non-specific adsorption.
  • CBMA1/BMA unit ratio
  • FIG. 1 schematically shows a sensor 2 of a measuring device 100 according to this embodiment.
  • the measurement device 100 can detect a specific substance (first substance) as a target from a measurement target (sample).
  • the first substance 5 is, for example, an in-vivo substance. Examples of the above-mentioned biological substances include proteins, DNA, substrates for enzymatic reactions, and the like.
  • the measuring device 100 includes a sensor 2 that can detect the first substance and a control device 6 that can control the measuring device 100.
  • the sensor 2 may be any sensor that uses, for example, elastic waves, QCM (Quartz Crystal Microbalance), SPR (Surface Plasmon Resonance), or FET (Field Effect Transistor). That is, the sensor 2 only needs to be able to mutually convert electrical signals and elastic waves, QCM, SPR, FET, etc.
  • the sensor 2 according to one embodiment is a sensor that uses elastic waves. That is, by using the sensor 2, the measuring device 100 according to one embodiment can detect a change in the elastic wave based on the presence of the first substance as a change in the electrical signal.
  • the sensor 2 may be manufactured by a conventionally known method, except for the polymer film 1 described below. In this case, the inspection information included in the identification information includes the initial phase of the elastic wave, the orientation of the substrate 22, etc. , information specific to a sensor that uses elastic waves may be included.
  • the sensor 2 has an external terminal 21.
  • the sensor 2 can be electrically connected to a control device 6 that controls the measuring device 100 via an external terminal 21 . That is, the sensor 2 and the control device 6 can input and output electrical signals to each other via the external terminal 21. Therefore, the control device 6 can detect, for example, the first substance based on the electrical signal input from the sensor 2. For example, the control device 6 may calculate the concentration of the first substance contained in the sample. Alternatively, the control device 6 may identify the first substance, for example.
  • the control device 6 and the external terminals 21 may be manufactured using conventionally known techniques. Further, the configuration for electrically connecting the sensor 2 and the control device 6 is not limited to the external terminal 21.
  • the sensor 2 and the control device 6 may be electrically connected by electromagnetic induction.
  • the sensor 2 may be a disposable cartridge. According to this, the step of cleaning the sensor 2 after measurement is unnecessary, and the influence of insufficient cleaning on the measurement results can be eliminated.
  • FIG. 2 shows a plan view of the sensor 2.
  • the sensor 2 includes a substrate 22, a detection section 23 located on the substrate 22, a reference section 24, and a pair of first IDT (Inter Digital Transducer) electrodes 25a arranged on the substrate 22 so as to sandwich the detection section 23 therebetween. , a pair of second IDT electrodes 25b.
  • the detection section 23, the reference section 24, the pair of first IDT electrodes 25a, and the pair of second IDT electrodes 25b may be located on the substrate 22.
  • the substrate 22 is, for example, a piezoelectric substrate.
  • the substrate 22 is, for example, a crystal substrate.
  • the substrate 22 is not limited to a crystal substrate as long as it can propagate elastic waves. That is, the substrate 22 may be made of any material that can propagate elastic waves.
  • the substrate 22 may be a substrate containing metals such as gold, silver, copper, platinum, and aluminum, lithium tantalate, and a piezoelectric single crystal such as quartz. Further, the substrate 22 may be manufactured by a conventionally known method.
  • a substance (second substance 4) that reacts with the first substance 5 is fixed to the detection unit 23. Therefore, in the detection unit 23, the first substance 5 contained in the specimen and the fixed second substance 4 can react.
  • the detection unit 23 changes the propagation characteristics of the elastic waves of the substrate 22 by causing the first substance 5 and the second substance 4 to react. Specifically, the detection unit 23 changes the weight applied to the substrate 22 or the viscosity of the liquid that contacts the surface of the substrate 22 by causing the first substance 5 and the second substance 4 to react, for example. The magnitude of these changes correlates with the amount of reaction between the first substance 5 and the second substance 4. Further, the characteristics of the elastic wave (for example, phase, amplitude, period, etc.) change as it propagates through the detection unit 23.
  • the magnitude of the change in characteristics correlates with, for example, the magnitude of the weight applied to the substrate 22 or the magnitude of the viscosity of the liquid that contacts the surface of the substrate 22. Therefore, the sensor 2 can detect the first substance 5 based on changes in the characteristics of the elastic waves. Specifically, the measuring device 100 can measure, for example, the concentration of the first substance 5 contained in the sample. Details of the detection unit 23 will be described later.
  • the pair of first IDT electrodes 25a can generate elastic waves between the pair of first IDT electrodes 25a.
  • the elastic waves that propagate on the surface of the substrate 22 are also referred to as surface acoustic waves (SAW).
  • SAW surface acoustic waves
  • the pair of first IDT electrodes 25a may be located on the substrate 22 so as to sandwich the detection section 23 therebetween.
  • an electrical signal is input to one of the pair of first IDT electrodes 25a.
  • the input electrical signal is converted into an elastic wave that propagates toward the detection section 23 and is emitted from one first IDT electrode 25a.
  • the emitted elastic waves pass through the detection section 23.
  • the other first IDT electrode 25a can receive the elastic wave that has passed through the detection section 23.
  • the received elastic waves are converted into electrical signals.
  • the pair of first IDT electrodes 25a may be made of, for example, a metal material such as gold, chromium, or titanium. Further, the pair of first IDT electrodes 25a may be a single layer electrode made of a single material, or a multilayer electrode made of a plurality of materials.
  • the sensor 2 may have two or more combinations of the detection section 23 and the pair of first IDT electrodes 25a.
  • the measuring device 100 may detect different types of target substances for each combination, for example.
  • the measuring device 100 may, for example, detect the same type of target substance in multiple combinations and compare the respective detection results.
  • FIG. 3 shows a conceptual diagram showing the polymer membrane 1 included in the detection section 23.
  • a polymer film 1 containing a polymer 3 is fixed on a substrate 22.
  • a substance (second substance 4) that reacts with the first substance 5 is fixed on the polymer film 1.
  • the polymer membrane 1 is a membrane that has been adjusted to have high specific adsorption and is a membrane that has been adjusted to reduce non-specific adsorption.
  • the reaction between the first substance 5 and the second substance 4 may be any reaction that causes a change in the output of the sensor 2.
  • a reaction may be, for example, a reaction in which the first substance 5 and the second substance 4 are bonded together through a redox reaction, an enzyme reaction, an antigen-antibody reaction, chemisorption, intermolecular interaction, or ionic interaction. It's okay.
  • the reaction between the first substance 5 and the second substance 4 may be a reaction in which a new substance (third substance) is produced by an enzymatic reaction or the like.
  • the second substance 4 to be fixed to the detection unit 23 may be appropriately selected depending on the first substance 5.
  • the second substance 4 may be an antibody, a peptide, an aptamer, or the like.
  • the second substance 4 may be an antigen.
  • the second substance 4 may be an enzyme.
  • the measuring device 100 may indirectly detect the first substance 5 that is the target.
  • a substance similar to the first substance 5 may be fixed to the detection unit 23 as the second substance 4. That is, for example, an antibody whose antigen is the first substance 5 may be reacted with the first substance 5 in advance, and the unreacted antibody may be reacted with the immobilized second substance 4.
  • the measuring device 100 can indirectly calculate the amount of the first substance 5 from the amount of detected antibody.
  • the triblock copolymer of the present disclosure is a copolymer containing two hydrophilic blocks A and a hydrophobic block B between the two hydrophilic blocks.
  • Hydrophilic block A includes at least one structural unit (a) among the structural units (a) represented by the following formulas (1) to (6).
  • R 1 to R 7 are each independently H or CH 3 .
  • X 1 to X 4 are each independently O or NH
  • p 1 to p 5 are each independently an integer of 1 or more and 3 or less (1, 2 or 3).
  • q 1 to q 2 are each independently an integer from 1 to 5 (1, 2, 3, 4, or 5).
  • the structural unit (a) may have a betaine structure.
  • the structural unit (a) may have a COO 2 - group in terms of making it easier to immobilize antibodies and the like.
  • Examples of the structural unit (a) include the following structural units.
  • the proportion of the structural unit (a) constituting the hydrophilic block A contained in the triblock copolymer of the present disclosure may be 20 mol% or more, and may be 30 mol% or more in terms of reducing nonspecific adsorption.
  • the amount may be 40 mol% or more.
  • the above ratio may be 70 mol% or less, 60 mol% or less, or 50 mol% or less.
  • the hydrophilic block A may have a structural unit other than the structural unit (a) within a range that does not impair the effects of the present disclosure.
  • each structural unit may be contained in the block A by any mode such as random copolymerization or block copolymerization.
  • the two hydrophilic blocks A contained in the triblock copolymer of the present disclosure may have the same or different structures. In the case where two hydrophilic blocks A have the same structure, it is sufficient that they have the same structural unit (a) within a range that does not impair the effects of the present disclosure.
  • the equivalent structural unit (a) may have the same structural unit (a), and may have a structural unit other than the structural unit (a).
  • the equivalent structural unit (a) may have the same structure as the structural unit (a), for example, the relative position and number average degree of polymerization of the structural unit (a) may be the same. .
  • Hydrophobic block B includes at least one structural unit (b) among the structural units (b) represented by the following formulas (7) and (8).
  • R 8 and R 10 are each independently H or CH 3 .
  • R 9 , R 11 and R 12 are each independently an alkyl group having 1 or more and 6 or less carbon atoms.
  • X 5 and X 6 are O or NH.
  • r is an integer (1, 2 or 3) of 1 or more and 3 or less.
  • alkyl group having 1 to 6 carbon atoms examples include methyl group, ethyl group, propyl group, isopropyl group, butyl group, isobutyl group, sec-butyl group, tert-butyl group, heptyl group, hexyl group, etc. .
  • Examples of the structural unit (b) include the following structural units.
  • the proportion of the structural unit (b) constituting the hydrophobic block B contained in the triblock copolymer of the present disclosure may be 30 mol% or more in terms of ease of forming micelles of the polymer. , may be 40 mol% or more, or may be 50 mol% or more. Further, the above ratio may be 80 mol% or less, 60 mol% or less, or 50 mol% or less.
  • the hydrophobic block B may have a structural unit other than the structural unit (b) as long as the effects of the present disclosure are not impaired.
  • each structural unit may be contained in the block B by any mode such as random copolymerization or block copolymerization.
  • Hydrophobic block B has a disulfide bond between structural units (b).
  • the number of disulfide bonds may be one or more.
  • At least one end of the triblock copolymer of the present disclosure may have a thiol group or a dithioester group from the viewpoint of ease of fixing the polymer membrane to the substrate 22 (measurement substrate 12). , may be a thiol group.
  • the ratio of the number of moles of the structural unit (a) to the number of moles of the structural unit (b) contained in the triblock copolymer of the present disclosure is determined to reduce nonspecific adsorption and form micelles of the copolymer. In terms of ease, it may be 0.20 or more, 0.50 or more, or 0.70 or more. Further, the above ratio may be 2.5 or less, 2.0 or less, or 1.5 or less.
  • the number average degree of polymerization of the triblock copolymer of the present disclosure may be 100 or more, or even 200 or more, in terms of ease of forming micelles of the copolymer, film formation density, etc. good.
  • the total number average degree of polymerization of the two hydrophilic blocks A in the triblock copolymer of the present disclosure may be 30 or more, 60 or more, 100 or more in terms of reducing nonspecific adsorption. It may be 200 or less, or it may be 150 or less.
  • the number average degree of polymerization of the hydrophobic blocks B in the triblock copolymer of the present disclosure may independently be 25 or more, and 30 It may be more than 50, it may be more than 80.
  • the triblock copolymer of the present disclosure may be identified by conventionally known organic analysis techniques.
  • the copolymer may be identified by NMR (Nuclear Magnetic Resonance).
  • NMR Nuclear Magnetic Resonance
  • it may be identified, for example, by liquid chromatography.
  • infrared spectroscopy That is, when identifying the copolymer, an apparatus capable of implementing these techniques may be used.
  • the identification method and device are not limited to these methods and devices.
  • An example of the method for producing a triblock copolymer of the present disclosure includes a step of synthesizing a hydrophobic block B (step 1) and a step of synthesizing a hydrophilic block A (step 2). Each step will be explained below.
  • hydrophobic block B is synthesized by polymerizing a hydrophobic monomer compound.
  • hydrophobic monomer compounds include the following compounds.
  • butyl methacrylic acid (BMA) etc. are mentioned as a specific example of a hydrophobic monomer compound.
  • BMA butyl methacrylic acid
  • the hydrophobic monomer compounds may be used alone or in combination of two or more.
  • R 8 to R 12 , X 5 and X 6 and r have the same meanings as R 8 to R 12 , X 5 and X 6 in formulas (7) to (8), respectively.
  • Polymerization can be produced by known polymerization such as living radical polymerization (LRP).
  • living radical polymerization examples include atom transfer radical polymerization (ATRP), single electron transfer polymerization (SET-LRP), reversible chain transfer catalyzed polymerization (RTCP), and RAFT polymerization.
  • ATRP atom transfer radical polymerization
  • SET-LRP single electron transfer polymerization
  • RTCP reversible chain transfer catalyzed polymerization
  • RAFT polymerization Reversible Addition-Fragmentation chain Transfer Polymerization
  • polymerization initiators include polymerization initiators containing disulfide bonds such as bis[2-(2'-bromoisobutyryloxy)ethyl]disulfide (BiBOEDS)).
  • Step 2 Synthesis step of hydrophilic block A>
  • hydrophilic blocks A are synthesized at both ends of hydrophobic block B by polymerizing the polymer obtained in step 1 with a hydrophilic monomer compound, thereby obtaining the triblock copolymer of the present disclosure.
  • hydrophilic monomer compounds include the following compounds (11) to (16).
  • hydrophilic monomer compound N-(carboxymethyl)-N,N-dimethyl-2-[(2-methyl-1-oxo-2-propen-1-yl)-oxy]ethanaminium (CBMA1 ), N,N-dimethylaminoethyl methacrylate (DMAEMA), 2-hydroxypropyl methacrylamide (HPMA), and the like.
  • the hydrophilic monomer compounds may be used alone or in combination of two or more. (In the formula, R 1 to R 7 , X 1 to X 4 , p 1 to p 5 and q 1 to q 2 are R 1 to R 7 , X 1 to 4 , p 1 to p 5 and q 1 to q 2. )
  • the method for producing a triblock copolymer of the present disclosure may include a quaternary ammonium salt formation step (Step 3) and a deprotection step (Step 4).
  • Step 3 Quaternary ammonium formation step of hydrophilic block A>
  • the tertiary amine structure of the structural unit (a) of the hydrophilic block A of the polymer obtained in Step 2 is converted into a quaternary ammonium structure (quaternary ammonium formation).
  • quaternary ammonium formation may be performed using a known quaternizing agent such as a halogen compound.
  • Betainization step of hydrophilic block A> a betaine structure is formed in the structural unit (a) of the hydrophilic block A of the polymer obtained in step 3 (betaine formation).
  • betaination may be performed by deprotecting the side chain protecting group (eg, tert-butyl group) of the hydrophilic block A introduced in step 3.
  • the structure derived from the polymerization initiator in the produced triblock copolymer may be removed.
  • An example of a structure derived from the polymerization initiator is bromine at the terminal of a triblock copolymer when LRP is performed using BiBOEDS as a polymerization initiator.
  • the polymer membrane of the present disclosure includes a copolymer containing a hydrophilic block A and a hydrophobic block B.
  • the copolymer is a diblock copolymer, and the diblock copolymer is also included in one embodiment of the present disclosure.
  • the diblock copolymer of the present disclosure corresponds to polymer 3 in FIG.
  • the end of the hydrophobic block B opposite to the side bonded to the hydrophilic block A has a thiol group or a dithioester group.
  • the structural units of the hydrophilic block A and the hydrophobic block B of the diblock copolymer of the present disclosure and their specific examples are the structural units of the hydrophilic block A and the hydrophobic block B of the triblock copolymer of the present disclosure and their specific examples. This is the same as the specific example.
  • the ratio of the number of moles of the structural unit (a) to the number of moles of the structural unit (b) contained in the diblock copolymer of the present disclosure is 0.20 in terms of nonspecific adsorption reduction and film formation density. or more, may be 0.50 or more, or may be 0.70 or more. Further, the above ratio may be 2.5 or less, 2.0 or less, or 1.5 or less.
  • the number average degree of polymerization of the diblock copolymer of the present disclosure may be 25 or more, 50 or more, or 200 or more in terms of film forming density.
  • the diblock copolymer of the present disclosure may have a high molecular weight.
  • the number average degree of polymerization of the hydrophilic block A in the diblock copolymer of the present disclosure may be 20 or more, 40 or more, or 60 or more in terms of reducing nonspecific adsorption. Generally, it may be 80 or more, 100 or more, or 200 or less.
  • the number average degree of polymerization of the hydrophobic blocks B in the diblock copolymer of the present disclosure may independently be 25 or more, 30 or more, or 50 or more in terms of film forming density.
  • the number may be 80 or more.
  • the diblock copolymer of the present disclosure may be identified by conventionally known organic analysis techniques.
  • the diblock copolymer of the present disclosure can be obtained by cleaving the disulfide bonds of the triblock copolymer of the present disclosure or micelles of the copolymer.
  • a conventionally known reducing agent may be used to cleave the disulfide bond.
  • An example of the method for producing a polymer film of the present disclosure includes a step of preparing a coating agent containing micelles of the triblock copolymer of the present disclosure, a step of disposing the coating agent on a substrate, and a step of preparing a coating agent containing micelles of the triblock copolymer of the present disclosure. cleaving disulfide bonds in micelles of the polymer to form a diblock copolymer.
  • Micelles of the triblock copolymer of the present disclosure are also included in one embodiment of the present disclosure.
  • the micelles are flower-shaped micelles.
  • an example of the method for producing a polymer membrane of the present disclosure includes a step of preparing a coating agent containing micelles of a diblock copolymer obtained by cleaving the disulfide bonds of the triblock copolymer of the present disclosure; arranging the coating agent on the substrate.
  • Micelles of diblock copolymers obtained by cleaving the disulfide bonds of the triblock copolymers of the present disclosure are also included in one embodiment of the present disclosure.
  • the micelles are W/O type micelles.
  • a polymer membrane formed by a diblock copolymer obtained by cleaving disulfide bonds after forming micelles of the triblock copolymer of the present disclosure has a high density of the diblock copolymer. Furthermore, a high density polymer membrane can also be achieved by using micelles of a diblock copolymer obtained by cleaving the disulfide bonds of the triblock copolymer of the present disclosure. Furthermore, the polymer membrane formed by the diblock copolymer obtained from the triblock copolymer of the present disclosure has a reduced amount of nonspecific adsorption.
  • Methods for immobilizing the diblock copolymer of the present disclosure on the substrate 22 include, for example, a method in which a polymer solution obtained by dissolving polymer 3 in a solvent is applied to the substrate 22 and dried, graft polymerization using radiation or ultraviolet rays, Examples include a chemical reaction with a functional group of the substrate 22. By these methods, the polymer film 1 made of the polymer 3 is formed on the substrate 22.
  • An example of a method for fixing the second substance 4 onto the polymer 3 (polymer membrane 1) is a method of covalently bonding the second substance 4 to a carboxyl group that the polymer 3 has.
  • polymer 3 is reacted with N-hydroxysuccinimide (NHS) and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) (NHS/EDC activation).
  • NHS N-hydroxysuccinimide
  • EDC 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide
  • NHS/EDC activation of the polymer 3 may be performed before immobilizing on the substrate 22 or after immobilizing the substrate 22. Since the polymer of this embodiment has high chemical stability, it is unlikely to be decomposed by NHS/EDC activation. Therefore, non-specific adsorption caused by decomposition products accompanying NHS/EDC activation can be reduced.
  • FIG. 4 shows a schematic configuration of the measurement carrier 11 according to this embodiment.
  • An example of the measurement carrier 11 is a plate for ELISA (enzyme-linked immunosorbent).
  • the measurement carrier 11 has a detection region 31 for specifically capturing the target substance (first substance 5) contained in the sample and a non-detection region 32 for non-selectively adsorbing a blocking agent etc. on the surface of the measurement substrate 12.
  • a detection region 31 for specifically capturing the target substance (first substance 5) contained in the sample
  • a non-detection region 32 for non-selectively adsorbing a blocking agent etc. on the surface of the measurement substrate 12.
  • We are preparing for The polymer film 1 is fixed to the detection region 31 .
  • a second substance 4 that reacts with the first substance 5 is fixed on the polymer membrane 1, similar to the polymer membrane 1 described in the first embodiment.
  • a membrane adjusted to increase non-specific adsorption may be fixed to the non-detection region 32.
  • a desired second substance 4 is fixed on the polymer film 1 in the detection region 31 . Further, the blocking agent is non-selectively adsorbed to the non-detection region 32 . Thereafter, by bringing the specimen into contact with the detection region 31, the first substance 5, which is a target substance contained in the specimen, reacts with the second substance 4, and the reaction is detected by the detection reagent.
  • detection reagents include redox substances, fluorescent substances, enzymes, and dye compounds.
  • the measurement substrate 12 may be made of, for example, metals such as gold, silver, copper, platinum, and aluminum; plastics such as polyethylene and polypropylene; and inorganic materials such as titanium oxide, silica, glass, and ceramics.
  • the measurement substrate 12 is not limited to these examples.
  • the shape of the measurement substrate 12 may be, for example, a plate, a particle, a microstructure, a microtiter plate, or the like.
  • the shape of the measurement substrate 12 is not limited to these examples.
  • a measurement kit including a measurement substrate 12 on which a polymer film 1 is fixed, a second substance 4, and a detection reagent is also included within the scope of the present disclosure.
  • the second substance 4 may be fixed to the polymer membrane 1 in advance during product manufacture, or may be fixed by the user before measurement.
  • the measurement kit according to this embodiment may include other reagents and instruments.
  • components other than the second substance 4 and the detection reagent described above may be included.
  • a buffer or the like may be provided.
  • the measurement kit according to the present embodiment may include a plurality of different reagents mixed in appropriate volumes and/or forms, or may be provided in separate containers.
  • the measurement kit according to the present embodiment may include an instruction sheet describing the procedure for detecting the reaction between the first substance 5 and the second substance 4. It may be written or printed on paper or other media, or it may be attached to an electronic medium such as a magnetic tape, a computer readable disk, or a CD-ROM or the like.
  • the copolymer according to aspect 1 of the present disclosure comprises two hydrophilic blocks A containing at least one structural unit (a) among the structural units (a) represented by the following formulas (1) to (6).
  • R 1 to R 7 are each independently H or CH 3
  • X 1 to X 4 are each independently O or NH
  • p 1 to p 5 are each independently 1 to 3
  • q 1 to q 2 are each independently an integer between 1 and 5.
  • a hydrophobic block B containing at least one structural unit (b) among the structural units (b) represented by the following formulas (7) and (8)
  • R 8 and R 10 are each independently H or CH 3
  • R 9 , R 11 and R 12 are each independently an alkyl group having 1 to 6 carbon atoms
  • X 5 and 6 is O or NH
  • r is an integer from 1 to 3.
  • the hydrophobic block B has a disulfide
  • the proportion of the structural unit (b) constituting the hydrophobic block B contained in the copolymer is 30 mol% or more and 80 mol% or less. It may be.
  • the proportion of the structural unit (a) constituting the hydrophilic block A contained in the copolymer is 20 mol % or more and 70 mol %. % or less.
  • the copolymer according to Aspect 4 of the present disclosure is provided in any one of Aspects 1 to 3, in which the number of moles of the structural unit (a) is 1 per mole of the structural unit (b) contained in the copolymer.
  • the numerical ratio may be 0.20 or more and 2.5 or less.
  • At least one terminal of the copolymer according to any one of Aspects 1 to 4 may have a thiol group or a dithioester group.
  • the polymer membrane according to aspect 6 of the present disclosure includes a hydrophilic block A containing at least one structural unit (a) among structural units (a) represented by the following formulas (1) to (6); (In the formula, R 1 to R 7 are each independently H or CH 3 , X 1 to X 4 are each independently O or NH, and p 1 to p 5 are each independently 1 to 3 The following integers are shown, and q 1 to q 2 are each independently an integer between 1 and 5.) A hydrophobic block B containing at least one structural unit (b) among the structural units (b) represented by the following formulas (7) and (8), (In the formula, R 8 and R 10 are each independently H or CH 3 , R 9 , R 11 and R 12 are each independently an alkyl group having 1 to 6 carbon atoms, and X 5 and 6 is O or NH, and r is an integer from 1 to 3.)
  • the end of the hydrophobic block B opposite to the side bonded to the hydrophilic block A contains a copolymer
  • a measuring device includes the polymer membrane of Aspect 6 above.
  • a measurement carrier according to Aspect 8 of the present invention includes the polymer membrane of Aspect 6 above.
  • % represents mass %.
  • p(BMA) was synthesized by the SET-LRP method.
  • 2.08 mol/L of monomer butyl methacrylic acid (BMA) and 20.0 mol/L of bis[2-(2'-bromoisobutyryloxy)ethyl] disulfide (BiBOEDS) were added to 2-propanol that had been previously subjected to deoxygenation treatment.
  • 8 mmol/L, copper (II) bromide was dissolved at 2.08 mmol/L, and N,N,N',N'',N''-pentamethyldiethylenetriamine (PMDETA) was dissolved at 15.0 mmol/L. .
  • the obtained solution was added to a copper wire that had been reduced with hydrazine, and reacted in nitrogen at 40° C. for 6 hours.
  • the amount of copper wire was such that the concentration would be 305 mmol/L assuming that it was all dissolved.
  • the reaction solution was filtered and poured into a large amount of methanol (15 to 20 times the volume of the polymerization solution).
  • p(BMA) was obtained by filtering and drying the precipitate.
  • the degree of polymerization of the obtained p(BMA) was calculated by nuclear magnetic resonance (NMR) spectroscopy.
  • the area of the peak (peak around 2.9 ppm) derived from the methylene group bonded to sulfur of BiBOEDS in the 1 H-NMR spectrum and the side chain (methylene group bonded to oxygen) in the BMA unit of the polymer are determined. ) and the area of the peak (peak around 3.90 ppm to 4.00 ppm).
  • the degree of polymerization of the obtained p(BMA) was estimated to be 52.
  • DMAEMA N,N-dimethylaminoethyl methacrylate
  • DMAEMA Polymerization of DMAEMA was performed by the SET-LRP method using p(BMA) synthesized in Step 1 as a macroinitiator.
  • PMDETA was dissolved to a concentration of 10.8 mmol/L.
  • the obtained solution was added to a copper wire that had been reduced with hydrazine, and reacted in nitrogen at 40° C. for 8 hours.
  • the amount of copper wire was such that the concentration would be 220 mmol/L assuming that all of the copper wire was dissolved.
  • 45°C warm water was added.
  • the precipitate was filtered and dried to obtain p(BMA)-bp(DMAEMA).
  • the degree of polymerization of the obtained p(BMA)-bp(DMAEMA) was calculated by nuclear magnetic resonance (NMR) spectroscopy. Specifically, it was calculated from the ratio of the area of the peak around 1.5 ppm originating from the BMA unit and the area of the peak around 2.3 ppm originating from the DMAEMA unit of the polymer in the 1 H-NMR spectrum. As a result, the ratio of BMA units to DMAEMA units (BMA:DMAEMA) was estimated to be 52:42.
  • Betaining DMAEMA unit The side chain of the DMAEMA unit was betaineated according to the scheme below.
  • the block copolymer obtained in step 3 in which the side chain of the DMAEMA unit was quaternary ammonium was dissolved in trifluoroacetic acid and reacted at room temperature for 20 hours. After the reaction, trifluoroacetic acid was distilled off by evaporation, and polymer powder was recovered. A polymer powder was obtained by dissolving the polymer powder in methanol, adding diethyl ether to cause reprecipitation, and filtering and drying the precipitate.
  • Nuclear magnetic resonance (NMR) spectroscopy revealed that the peak derived from the tert-butyl group had disappeared in the 1 H-NMR spectrum. From the results of NMR spectroscopy, it was confirmed that the tert-butyl group was deprotected and the desired triblock copolymer p(BMA)-bp(CBMA1) was obtained.
  • the triblock copolymer obtained by the above synthesis scheme is represented by the following formula.
  • p(CBMA1)-p(BMA)-SS-p(BMA)-p(CBMA1) is the hydrophilic block A
  • p(BMA)-SS-p(BMA) is the hydrophobic block B
  • "SS" in hydrophobic block B represents a disulfide bond.
  • the degree of polymerization of each block of the triblock copolymer obtained by the above synthesis scheme was 42-52-SS-52-42. That is, by the above synthesis scheme, an ABA type triblock copolymer containing a hydrophobic block B containing a disulfide bond and having a degree of polymerization of 104 between two hydrophilic blocks A having a degree of polymerization of 42 was obtained. It was done.
  • a triblock copolymer (p(CBMA1)-p(BMA)-SS-p(BMA)-p(CBMA1)) having the following degree of polymerization was also produced. ⁇ 24-27-SS-27-24 ⁇ 24-31-SS-31-24 ⁇ 20-84-SS-84-20 ⁇ 71-31-SS-31-71 ⁇ 80-84-SS-84-80
  • the polymerization of HPMA was carried out by the SET-LRP method using p(BMA) in which the p(BMA) unit was an 86-mer synthesized in the same manner as in Step 1 as a macroinitiator.
  • Me 6 TREN tris[2-(dimethylamino)ethylamine
  • the obtained solution was added to a copper wire that had been reduced with hydrazine, and reacted in nitrogen at 40° C. for 91 hours.
  • the amount of copper wire was such that the concentration would be 305 mmol/L assuming that it was all dissolved.
  • water was added to the soluble portion of the solution.
  • the soluble portion was removed by a decanting method, and p(BMA)-bp(HPMA) was obtained by drying.
  • the degree of polymerization of the obtained p(BMA)-bp(HPMA) was calculated by nuclear magnetic resonance (NMR) spectroscopy.
  • Example 2 Measurement of non-specific adsorption amount The non-specific adsorption amount of fetal bovine serum to the polymer membrane was measured under the following conditions. Measuring device: Biacore X100 manufactured by GE Healthcare Measurement conditions: Running buffer: HBS-P Temperature: 25°C Flow rate: 10 ⁇ L/min Sample contact time: 9 minutes (sample injection volume 90 ⁇ L)
  • SIA kit Au manufactured by GE Healthcare
  • piranha solution was washed with piranha solution.
  • piranha solution was immersed in a 1 mmol/L ethanol solution of 1-dodecanethiol and left overnight.
  • the SPR chip was washed with ethanol and ultrapure water.
  • it was immersed for 18 hours in a methanol solution in which the triblock copolymer was dissolved at a concentration of 0.3 mg/mL to form a polymer film.
  • the chip was washed with ultrapure water and dried in a nitrogen stream to obtain an SPR chip with a polymer film formed thereon.
  • FIG. 5 is a graph showing the relationship between the unit ratio (CBMA1/BMA) of the triblock copolymer and the amount of non-specific adsorption.
  • FIG. 6 is a graph showing the relationship between the average degree of polymerization of CBMA1 in the triblock copolymer and the amount of non-specific adsorption.
  • the non-specific adsorption amount of the polymer membrane is preferably 20 ng/cm 2 or less, and it is known that the higher the film formation density, the lower the non-specific adsorption amount. From the results in Table 1 and Figures 5 and 6, when the unit ratio (CBMA1/BMA) of the triblock copolymer is 0.20 or more and 2.5 or less, the amount of nonspecific adsorption decreases, and the unit ratio is 1 or less. was found to be more preferable. Further, it was found that the number average degree of polymerization of CBMA1 is preferably up to about 70, and more preferably about 20 to 40. Since CBMA1 has the ability to suppress non-specific adsorption, at first glance it seems preferable to have more CBMA1 units. On the other hand, since it is thought that it is more advantageous to have fewer CBMA units for micelle formation, the results of this example suggest that forming micelles is effective in reducing non-specific adsorption.
  • the present disclosure can be used in a measurement device and a measurement plate that include a detection section on which a polymer film is formed.

Landscapes

  • Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Hematology (AREA)
  • Molecular Biology (AREA)
  • Urology & Nephrology (AREA)
  • Medicinal Chemistry (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Physics & Mathematics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Microbiology (AREA)
  • Organic Chemistry (AREA)
  • Polymers & Plastics (AREA)
  • Food Science & Technology (AREA)
  • Cell Biology (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Addition Polymer Or Copolymer, Post-Treatments, Or Chemical Modifications (AREA)

Abstract

Ce copolymère comprend : deux blocs A comprenant au moins un motif structural (a) choisi parmi les motifs structuraux (a) représentés par les formules (1) à (6) ; et un bloc B interposé entre les deux blocs A, le bloc B comprenant au moins un motif structural (b) choisi parmi les motifs structuraux (b) représentés par les formules (7) et (8). Le bloc B a une liaison disulfure entre les unités structurales (b).
PCT/JP2023/016392 2022-04-26 2023-04-26 Copolymère, film polymère, dispositif de mesure et support de mesure Ceased WO2023210671A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP2024517362A JPWO2023210671A1 (fr) 2022-04-26 2023-04-26

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2022072657 2022-04-26
JP2022-072657 2022-04-26

Publications (1)

Publication Number Publication Date
WO2023210671A1 true WO2023210671A1 (fr) 2023-11-02

Family

ID=88519069

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2023/016392 Ceased WO2023210671A1 (fr) 2022-04-26 2023-04-26 Copolymère, film polymère, dispositif de mesure et support de mesure

Country Status (2)

Country Link
JP (1) JPWO2023210671A1 (fr)
WO (1) WO2023210671A1 (fr)

Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2001504093A (ja) * 1996-10-11 2001-03-27 アルザ コーポレイション 融合性リポソーム組成物および方法
US20030181613A1 (en) * 2002-03-18 2003-09-25 Lele Bhalchandra Shripad Amphiphilic diblock, triblock and star-block copolymers and their pharmaceutical compositions
JP2004509181A (ja) * 2000-09-18 2004-03-25 ロディア・シミ ジスルフィド化合物の存在下における制御されたラジカル重合によるブロックポリマーの合成方法
WO2007063320A1 (fr) * 2005-12-03 2007-06-07 The University Of Sheffield Gelificateur polymere
JP2008506780A (ja) * 2004-07-19 2008-03-06 セレーター ファーマスーティカルズ、インク． 活性剤放出のための粒子状構築物
CN101563377A (zh) * 2006-08-17 2009-10-21 罗地亚运作公司 嵌段共聚物、其制备方法及在乳液中的用途
US20120245244A1 (en) * 2007-10-17 2012-09-27 Sankaran Thayumanavan Cleavable block copolymers, functionalized nanoporous thin films and related methods of preparation
CN104072694A (zh) * 2014-06-20 2014-10-01 北京科技大学 一种四重响应性嵌段胶束的制备方法及其应用
CN113289015A (zh) * 2021-05-13 2021-08-24 华中科技大学 调节光敏剂聚集程度的方法、纳米配位聚合物及其制备方法和应用

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115957153B (zh) * 2022-12-12 2025-07-29 华南理工大学 具有组织修复和消炎功效的纳米组合物及其制备方法和应用

Patent Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2001504093A (ja) * 1996-10-11 2001-03-27 アルザ コーポレイション 融合性リポソーム組成物および方法
JP2004509181A (ja) * 2000-09-18 2004-03-25 ロディア・シミ ジスルフィド化合物の存在下における制御されたラジカル重合によるブロックポリマーの合成方法
US20030181613A1 (en) * 2002-03-18 2003-09-25 Lele Bhalchandra Shripad Amphiphilic diblock, triblock and star-block copolymers and their pharmaceutical compositions
JP2008506780A (ja) * 2004-07-19 2008-03-06 セレーター ファーマスーティカルズ、インク． 活性剤放出のための粒子状構築物
WO2007063320A1 (fr) * 2005-12-03 2007-06-07 The University Of Sheffield Gelificateur polymere
CN101563377A (zh) * 2006-08-17 2009-10-21 罗地亚运作公司 嵌段共聚物、其制备方法及在乳液中的用途
US20120245244A1 (en) * 2007-10-17 2012-09-27 Sankaran Thayumanavan Cleavable block copolymers, functionalized nanoporous thin films and related methods of preparation
CN104072694A (zh) * 2014-06-20 2014-10-01 北京科技大学 一种四重响应性嵌段胶束的制备方法及其应用
CN113289015A (zh) * 2021-05-13 2021-08-24 华中科技大学 调节光敏剂聚集程度的方法、纳米配位聚合物及其制备方法和应用

Also Published As

Publication number Publication date
JPWO2023210671A1 (fr) 2023-11-02

Similar Documents

Publication Publication Date Title
US7943370B2 (en) Structure, target substance detection element and target substance detection kit
Brown et al. Synthesis of oligo (ethylene glycol) methacrylate polymer brushes
EP1208126B1 (fr) Brosses polymeres destinees a immobiliser des molecules sur une surface ou un substrat, dans lesquelles les polymeres presentent des segments dispersables ou solubles dans l'eau et capteurs y etant lies
US7205161B2 (en) Polymer brushes for immobilizing molecules to a surface or substrate having improved stability
US7728094B2 (en) Selfassembled grafted polymeric layer for use in biosensor technology
US8232344B2 (en) Structure, target substance-detecting element and target substance-detecting kit
JP7443396B2 (ja) 共重合体、測定装置および測定用担体
JP4630865B2 (ja) 物質固定化剤、それを用いた物質固定化方法及びそれを用いた物質固定化基体
JP2009139366A (ja) 生体分子固定化のための方法
US20120045772A1 (en) Affinity Hydrogel and Label Independent Detection Methods Thereof
WO2023210671A1 (fr) Copolymère, film polymère, dispositif de mesure et support de mesure
JP4818056B2 (ja) 標的物質を捕捉する捕捉分子を結合させるための結合用官能基を基体上に備えた構造体
JP7333462B2 (ja) 共重合体およびその製造方法、測定装置ならびに測定用担体
WO2007043498A1 (fr) Substrat dote d'un groupe fonctionnel de liaison
JP5672892B2 (ja) リガンド固定化用共重合体及び該共重合体によるリガンドの固定化方法
US20040018532A1 (en) Electronic sensor device
JP5345335B2 (ja) 新規双性イオン型多分岐樹脂、並びに、蛋白質チップ表面調節剤
EP1369692A2 (fr) Surface de capteur
JP2011007785A (ja) 標的分子相互作用物質固定化担体
JP2016020822A (ja) 分析用担体、その製造方法および使用方法
JP2006335912A (ja) 生理活性物質固定化剤
Brault Multifunctional Zwitterionic Surface Chemistry for Applications in Complex Media

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 23796411

Country of ref document: EP

Kind code of ref document: A1

WWE Wipo information: entry into national phase

Ref document number: 2024517362

Country of ref document: JP

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 23796411

Country of ref document: EP

Kind code of ref document: A1