EP3092068A1 - Hydrogel sensible pour mettre en évidence des biomolécules - Google Patents

Hydrogel sensible pour mettre en évidence des biomolécules

Info

Publication number
EP3092068A1
EP3092068A1 EP14815644.1A EP14815644A EP3092068A1 EP 3092068 A1 EP3092068 A1 EP 3092068A1 EP 14815644 A EP14815644 A EP 14815644A EP 3092068 A1 EP3092068 A1 EP 3092068A1
Authority
EP
European Patent Office
Prior art keywords
alkyl
responsive hydrogel
hydrogel
monomers
biomolecules
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.)
Withdrawn
Application number
EP14815644.1A
Other languages
German (de)
English (en)
Inventor
André LASCHEWSKY
Erik Wischerhoff
Martin Sütterlin
Jean-Philippe Couturier
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.)
Fraunhofer Gesellschaft zur Forderung der Angewandten Forschung eV
Original Assignee
Fraunhofer Gesellschaft zur Forderung der Angewandten Forschung eV
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 Fraunhofer Gesellschaft zur Forderung der Angewandten Forschung eV filed Critical Fraunhofer Gesellschaft zur Forderung der Angewandten Forschung eV
Publication of EP3092068A1 publication Critical patent/EP3092068A1/fr
Withdrawn legal-status Critical Current

Links

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
    • G01N33/53—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
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J13/00—Colloid chemistry, e.g. the production of colloidal materials or their solutions, not otherwise provided for; Making microcapsules or microballoons
    • B01J13/0052—Preparation of gels
    • B01J13/0065—Preparation of gels containing an organic phase
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J3/00—Processes of treating or compounding macromolecular substances
    • C08J3/02—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques
    • C08J3/03—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques in aqueous media
    • C08J3/075—Macromolecular gels
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J2300/00—Characterised by the use of unspecified polymers
    • C08J2300/14—Water soluble or water swellable polymers, e.g. aqueous gels
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J2333/00—Characterised by the use of homopolymers or copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and only one being terminated by only one carboxyl radical, or of salts, anhydrides, esters, amides, imides, or nitriles thereof; Derivatives of such polymers
    • C08J2333/04—Characterised by the use of homopolymers or copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and only one being terminated by only one carboxyl radical, or of salts, anhydrides, esters, amides, imides, or nitriles thereof; Derivatives of such polymers esters
    • C08J2333/06—Characterised by the use of homopolymers or copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and only one being terminated by only one carboxyl radical, or of salts, anhydrides, esters, amides, imides, or nitriles thereof; Derivatives of such polymers esters of esters containing only carbon, hydrogen, and oxygen, the oxygen atom being present only as part of the carboxyl radical

Definitions

  • the present invention relates to a hydrogel suitable for the detection of
  • Biomolecules can be used.
  • the detection step usually takes place via the use of a labeling reagent (eg a radiolabeled secondary antibody) or the species to be detected must itself be labeled (eg fluorescent labeling of DNA fragments).
  • a labeling reagent eg a radiolabeled secondary antibody
  • the species to be detected must itself be labeled (eg fluorescent labeling of DNA fragments).
  • biosensors eg. Based on surface plasmon resonance or interference phenomena that do not require labeling of the species to be detected. However, these require a complex instrumentation. It would be desirable to have direct optical detection of biological species without the use of labeling reagents and without additional instrumentation.
  • WO 03/025538 A2 describes a sensor for the determination of the concentration of a chemical species, wherein the sensor contains a periodic arrangement of colloidal particles in a hydrogel matrix.
  • molecular aggregates such as proteins, DNA or even viruses would be particularly desirable because of their high biological and medical relevance.
  • a responsive hydrogel which is chemically cross-linked, has a porous photonic crystal structure and contains biomolecule-specific recognition groups.
  • hydrogels in the form of a porous photonic crystal are accessible by template synthesis, initially introducing a photonic crystal of colloidal particles as a template and polymerizing a hydrogel in the interstices between these colloidal particles, followed by removal of the colloidal ones Particles to obtain a porous photonic crystal structure. It has surprisingly been found in the context of the present invention that the porosity of such a photonic crystal is sufficient to reduce the diffusion of larger molecules, in particular biomolecules such. of biooligomers,
  • Biopolymers or biological particles Since it was therefore found that the detection of biomolecules, in particular larger biomolecules such as bio-oligomers, with the hydrogel structure according to the invention,
  • the hydrogel biomo contains molecule-specific recognition groups. Furthermore, it has been shown in the context of the present invention that a porous photonic crystal by a Responsive hydrogel (ie a hydrogel, which can change its swelling behavior under the influence of an external stimulus) is formed, a color change even in the wavelength range of visible light and therefore can make a complex instrumentation superfluous.
  • a Responsive hydrogel ie a hydrogel, which can change its swelling behavior under the influence of an external stimulus
  • a hydrogel is a three-dimensional network that is no longer soluble in water, but absorbs it and swells with it.
  • the crosslinking of the polymer chains can take place physically or chemically.
  • the network nodes are formed by entanglements and entanglements of long polymer chains with each other.
  • network nodes via physical interactions such.
  • electrostatic interactions are formed.
  • the junctions are formed by covalent bonds between the polymer chains.
  • the hydrogel is chemically crosslinked. This ensures that the porous photonic crystal structure of the hydrogel has sufficient stability.
  • the hydrogel according to the invention is a responsive hydrogel.
  • responsive hydrogels those skilled in the art will understand those hydrogels which, under the influence of an external stimulus (i.e., under the influence of an external, changing parameter), swell or may collapse under fluid delivery, alternatively. These transitions are preferably reversible.
  • hydrogels are generally known to the person skilled in the art and are also referred to as “stimuli-responsive hydrogels” or “switchable hydrogels”.
  • responsive hydrogel for example, those materials are suitable which have a volume phase transition. This can be on a lower and / or based in an upper critical solution temperature, however, in the case of crosslinked systems, no dissolution of the polymer takes place in the solvent. In such a system, for example, the temperature may act as a switch. In this context is also used by thermoresponsive hydrogels
  • Ionic strength, light or chemical reactions serve as a switch. If the system is suitably designed, even complex biomacromolecules such as proteins can act as a stimulus. Such systems are described, for example, in J. Buller et al, Polym. Chem. 2, 1486-1489 (2011).
  • Examples of groups which cause switchability with light are azobenzenes (Kröger R. et al., Macromol Chem. Phys. (1994) 195, 2291-2298) or spiropyrans (Edahiro et al, Biomacromolecules (2005) 6, 970-974 ).
  • Examples of groups which cause switchability via the pH are amino or carboxyl groups.
  • An example of switchability via a chemical reaction is described in P. Mi et al., Macromol. Rapid Commun. (2008) 29, 27-32.
  • the responsive hydrogels are those selected by an external stimulus (ie, by changing an external parameter) selected from temperature, pH, ionic strength, ionic species, electromagnetic radiation (especially light), a chemical Reaction, the presence (addition or replacement) of chemical (especially low molecular weight) or biochemical reagents or exposure to biomolecules such as proteins, or combinations thereof, swell or shrink.
  • an external stimulus ie, by changing an external parameter
  • an external parameter selected from temperature, pH, ionic strength, ionic species, electromagnetic radiation (especially light), a chemical Reaction, the presence (addition or replacement) of chemical (especially low molecular weight) or biochemical reagents or exposure to biomolecules such as proteins, or combinations thereof, swell or shrink.
  • an external stimulus ie, by changing an external parameter
  • an external parameter selected from temperature, pH, ionic strength, ionic species, electromagnetic radiation (especially light), a chemical Reaction, the presence (addition or replacement) of chemical (especially low molecular weight
  • the responsive hydrogel of the present invention contains biomolecule-specific recognition groups. It is preferred that the hydrogel either swells or shrinks by binding the biomolecules to the specific recognition groups. In a preferred embodiment, the responsive hydrogel by binding the biomolecules to the specific recognition groups one
  • volume phase transition shows the responsive hydrogel under isothermal conditions as a result of binding of biomolecules and a consequent change in the hydrophilic or hydrophobic character, a sudden change in volume.
  • the volume change of the responsive hydrogel which is caused by the binding of the biomolecule to be detected to the specific recognition groups, changes the size of the unit cell of the photonic crystal and thus also the peak maximum of the Bragg reflection. This peak shift can be used for the analytical detection of biomolecules.
  • Suitable monomer units for responsive hydrogels are generally known to the person skilled in the art.
  • the responsive hydrogel contains one or more of the following monomer units (and optionally others
  • Ri H, alkyl such as. B. CI_ 4 alkyl, preferably -CH 3;
  • x 0-50, more preferably 1-50 or 1-20,
  • Ri H, alkyl such as. B. CI_ 4 alkyl, preferably -H;
  • x 0-50, more preferably 1-50 or 3-20, (3)
  • x 0-50, more preferably 1-50 or 2-20, (4)
  • x 0-50, more preferably 1-50 or 2-20,
  • Ri H, alkyl such as C 1-4 alkyl
  • Ri H, alkyl such as Ci_ 4 alkyl, preferably H;
  • x 1-6, more preferably 3-4,
  • R 2 alkyl such as CI_ 4 alkyl independently H;
  • Ri H or a branch of the polysaccharide chain
  • R 2, R 3, R 5, R E independently of one another H, alkyl such as CI_ 4 alkyl, allyl, - (CH 2) "- COOH or a salt thereof,
  • Polysaccharide units such as, for example, carboxymethylcellulose units, hydroxyethyl starch units,
  • Ri H, alkyl such as CI_ 4 alkyl preferably -CH 3,
  • Ri H, alkyl such as CI_ 4 alkyl preferably -CH 3,
  • the respective monomer units may be randomly distributed over the polymer chain or in block form. It is preferably a random copolymer.
  • the responsive hydrogel contains the following monomer units (Ia) and (Ib) (and optionally further monomer units for more precise fine tuning of the material properties):
  • Ri H, alkyl such as. B. CI_ 4 alkyl, preferably -CH 3;
  • Ri H, alkyl such as. B. CI_ 4 alkyl, preferably -CH 3,
  • Recognition groups have good accessibility for the biomolecules to be detected. If the biomolecules to be detected bind to the recognition groups, there is a significant volume change and thus also a very clear shift of the peak maximum of the Bragg reflection.
  • the proportion of monomer units (Ia) and (Ib) can vary over a wide range.
  • the monomer units (Ia) may be present in an amount of 30-90 mol%, more preferably 50-80 mol%, based on the total amount of the monomers.
  • the monomer units (Ib) may be present, for example, in an amount of 2-40 mol%, more preferably 5-35 mol%, based on the total amount of the monomers.
  • the monomer units (Ia) may have a reactive group in their side chain, in particular -OH, or -COOH.
  • the monomer units (Ia) have the following structure:
  • x 0 or 1 or 2.
  • the monomer units (Ia) may be a mixture of at least two different monomer units (i) and (ii), which have the following structures:
  • Ri H, alkyl such as. B. Ci_ 4 alkyl, preferably -CH 3
  • Ri H, alkyl such as. As Ci_ 4 alkyl, preferred
  • R 2 alkyl, such as. B. CI_ 4 alkyl
  • x 0 or 1 or 2.
  • the responsive hydrogel of the present invention is chemically crosslinked.
  • the skilled worker is generally aware of how hydrogels can be chemically crosslinked.
  • chemical crosslinking is effected by making the hydrogel in the presence of crosslinkable monomers, and preferably, the crosslinkable monomers are present in an amount of from 2 to 20 mole percent, based on the total amount of the monomers.
  • this amount of crosslinking monomers has proved to be advantageous that the hydrogel and thus the porous photonic crystal structure have sufficient stability, on the other hand, the specific recognition groups for the biomolecules to be detected are still easily accessible.
  • crosslinkable monomers may be photocrosslinkable or thermally crosslinkable monomers. Suitable monomers for this purpose are known in principle to the person skilled in the art.
  • the crosslinkable monomers can be, for example, multifunctional (for example bi-, tri- or tetra-functional) monomers, ie monomers having two or more
  • Suitable multifunctional monomers are e.g. multifunctional acrylic, methacrylic, vinyl or allyl monomers.
  • di (meth) acrylates or tri (meth) acrylates which may optionally also be ethoxylated, can be used as multifunctional crosslinker monomers.
  • ethoxylated di (meth) acrylates may have the following chemical formula:
  • Ri and R 2 are independently H or methyl and
  • n 1-5000, more preferably 1-100 or 1-30.
  • Suitable photocrosslinkable monomers i.e., monomers which have a
  • Photochemical reaction the crosslinking of adjacent polymer chains cause
  • photocrosslinkable monomers contain a photoreactive group, for example a benzophenone group, an acetophenone group, a diazirine group or an azide group.
  • the responsive hydrogel of the present invention has a porous photonic crystal structure.
  • the term "photonic crystal” is used in its usual meaning known to the person skilled in the art and therefore refers to a material with a spatially periodically varying refractive index, the period length being comparable to the wavelength of the light and the material being a Bragg
  • the material itself, which forms the photonic crystal does not have to be crystalline, it is crucial that the material (in the case of the present invention, the responsive hydrogel) is spatially arranged to result in a periodically varying refractive index If this period length changes, for example because the photonic crystal shrinks or swells, the peak position of the Bragg reflection also changes If the shift in the peak positions of the Bragg reflection occurs in the visible wavelength range of the light, the extent of the Bragg reflection is
  • Opals are a well-known example of a photonic crystal or a material with a photonic crystal structure. Due to the porosity of the photonic formed by the hydrogel
  • Crystal structure is a diffusion of larger biomolecules in this structure readily possible and thus all specific recognition groups of the hydrogel are in principle accessible.
  • such a porous photonic crystal structure is obtained by first preparing a photonic crystal of colloidal particles, which are preferably monodisperse, and forming a chemically cross-linked hydrogel in the interstices of these colloidal particles.
  • the colloidal particles are preferably packed so densely that one particle touches as many of its neighboring particles as possible. Preferably, therefore, the closest possible packing of the colloidal particles is present. No polymer is formed at these contact surfaces of adjacent particles.
  • the colloidal particles are removed again, e.g. by a suitable solvent, leaving a porous photonic crystal structure.
  • the crystal structure of the hydrogel according to the invention is therefore obtained by a template-induced preparation process, wherein a photonic crystal of colloidal particles acts as a template.
  • the porous photonic crystal structure of the hydrogel is thus to a certain extent the negative of the structure of the photonic template crystal.
  • the porous photonic crystal structure is an inverse opal structure.
  • the diameter of the cavities of the porous photonic crystal structure can be controlled by the size of the colloidal particles of the photonic template crystal. These particles are preferably monodisperse particles. Preferably, the colloidal particles of the photonic template crystal have a coefficient of variation of ⁇ 20%, more preferably ⁇ 10% or even ⁇ 5%. Suitable colloidal particles for the production of a photonic crystal are commercially available or can also be obtained by conventional production methods known to the person skilled in the art.
  • the mean diameter of the colloidal, preferably monodisperse, particles of the photonic template crystal and thus also of the cavities of the porous photonic crystal structure can be varied over a wide range.
  • the average diameter may be, for example, in the range of 600 to 100 nm, more preferably 500 to 150 nm (for example, determined by
  • Wavelength range can be obtained. This in turn allows analytical detection of biomolecules in the wavelength range of visible light.
  • the cavities generated by such a template synthesis are interconnected and the passages between these cavities are sufficiently large to allow the detection of larger biomolecules.
  • biomolecule-specific recognition groups are those which are suitable for the detection of Biooligomeren, biopolymers or biological particles are suitable.
  • biomolecule-specific recognition groups used in this specification
  • antibodies F ab fragments of antibodies, enzymes, enzyme fragments, coenzymes, peptides, prosthetic groups, aptamers, single-stranded DNA and RNA single strands.
  • bio-oligomers are oligopeptides, oligosaccharides, oligonucleotides.
  • biopolymers are polypeptides, proteins, polysaccharides,
  • Exemplary biological particles are viruses.
  • biomolecule-specific recognition groups are preferably bound to the hydrogel via a covalent bond.
  • the covalent attachment of the biomolecule-specific recognition groups can be realized in the hydrogel, starting with the polymerization
  • Monomer compounds are present which contain such a biomolecule-specific recognition group. Alternatively, it is also possible that in this
  • the present invention relates to a process for producing a chemically crosslinked, responsive hydrogel having a porous photonic crystal structure comprising:
  • Suitable colloidal particles for the formation of photonic crystals are known in principle to the person skilled in the art. Preferably, it is monodisperse particles.
  • the colloidal particles may, for example, a
  • inorganic particles for example Si0 2 particles
  • organic polymer particles can be used. These particles must be selected so that they can then be removed again, eg under the influence of a
  • colloidal, preferably monodisperse particles are obtainable by conventional methods known to the person skilled in the art or else commercially available.
  • a dispersion of colloidal particles is preferably applied to a substrate and the liquid dispersion medium is allowed to evaporate slowly.
  • the colloidal particles deposit on the substrate in a periodically uniform array to form the photonic template crystal.
  • the monomers include the following
  • H 2 C C (R i) -C (O) -O-CH 2 -CH 2 - [CH 2 -CH 2 -O] x -R 2
  • Ri H, alkyl such as. B. CI_ 4 alkyl, preferably -CH 3,
  • H 2 C C (R i) -C (O) -O-CH 2 -CH 2 - [CH 2 -CH 2 -O] x -R 2
  • Ri H, alkyl such as. B. CI_ 4 alkyl, preferably -CH 3,
  • the proportion of monomers (a1) and (a2) can vary over a wide range.
  • the monomers (a1) may be present in an amount of 30-90 mol%, more preferably 50-80 mol%, based on the total amount of the monomers.
  • the monomer units (a2) may be present, for example, in an amount of 2-40 mol%, more preferably 5-35 mol%, based on the total amount of the monomers.
  • the monomers (a1) have a reactive group in their side chain, in particular -OH, or -COOH.
  • Ri H, alkyl such as. B. CI_ 4 alkyl, preferably -CH 3,
  • x 0 or 1 or 2.
  • the monomers (a1) may be a mixture of at least two different monomers (a1) and (al2) which have the following structures:
  • H 2 C C (R i) -C (O) -O-CH 2 -CH 2 - [CH 2 -CH 2 -O] x -R 2
  • Ri H, alkyl such as. B. CI_ 4 alkyl, preferably -CH 3,
  • H 2 C C (R i) -C (O) -O-CH 2 -CH 2 - [CH 2 -CH 2 -O] x -R 2
  • Ri H, alkyl such as. B. CI_ 4 alkyl, preferably -CH 3,
  • R 2 , alkyl such as. B. CI_ 4 alkyl
  • the chemical crosslinking is effected by making the hydrogel in the presence of crosslinkable monomers, and preferably using the crosslinkable monomers in an amount of 2-20 mol% to the total amount of monomers.
  • crosslinkable monomers reference may be made to the above statements.
  • Suitable polymerization conditions for the conversion of the monomers to the hydrogel are known to the person skilled in the art.
  • the covalent bonding of the biomolecule-specific recognition groups in the hydrogel can be realized by the presence of monomer compounds which are already present during the polymerization
  • Biomo contain molecule-specific recognition group.
  • a monomer compound having an organic functional group eg, -OH or -COOH
  • these organic functional groups with a compound containing the biomolecule-specific recognition group, be reacted. This is known in principle to the person skilled in the art.
  • the porous photonic crystal structure can be obtained by customary methods known to the person skilled in the art.
  • the colloidal particles can be removed by a solvent.
  • suitable organic solvents can be used.
  • SiCV particles can be removed, for example, by hydrofluoric acid (HF).
  • the present invention relates to a device for the detection of biomolecules, comprising the above-described
  • Responsive hydrogel according to the invention with a porous photonic crystal structure.
  • properties of the responsive hydrogel reference may be made to the above statements.
  • biomolecules which are preferably detected with the device, reference may be made to the above statements.
  • the device according to the invention for the detection of biomolecules may also have further device elements which are customary for this type of device, for example a signal converter and / or an electrical amplifier.
  • Biomolecules to the detection groups in the wavelength range of visible light can show a significant shift in the peak position of the Bragg reflection, it is within the scope of the present invention also possible that the
  • the present invention relates to the use of the hydrogel described above for the detection of biomolecules.
  • the detection of biomolecules can be done isothermally.
  • the preparation of the hydrogels in the form of porous photonic crystals was carried out by a template method.
  • the template used in each case was a photonic crystal of monodisperse particles.
  • monodisperse silica particles with a diameter of 400 nm were prepared by the established "Stöber method" (described in W. Stoeber et al., J. Colloid Interface Sei., 1968, 26, 62-69) Silica particles were then deposited vertically on a microscope slide.
  • the ethanolic silica dispersion was adjusted to a concentration of 2% by weight by adding ethanol and water (medium: 80% by weight of EtOH, 20% by weight of ultrapure water). Subsequently, the dispersion was in a
  • the photonic template crystals thus obtained had a vertical layer thickness of about 5 ⁇ m and exhibited pronounced opalescence.
  • the coated slide was then covered with another slide and sealed the resulting shape on three sides. Over the open side, a solution with the monomers, Irgacure 2010 as a UV initiator (1.5 wt .-% relative to the monomers) and water and ethanol was injected as a solvent (total content of monomers and crosslinkers in the solution: 35 wt. -%).
  • the polymerization solution filled in the interparticle spaces after injection. Subsequently, the polymerization form was irradiated with UV light (emission maximum 365 nm, 400 W, Hoenle Co., type UVA Cube) and the hydrogel was crosslinked in this way.
  • OEGMA300 01igo (ethylene glycol) methyl ether methacrylate, average
  • OEGMA4 01igo (ethylene glycol) methyl ether methacrylate, average
  • MEO2MA di (ethylene glycol) methyl ether methacrylate
  • OEGDMA 400 01igo (ethylene glycol) dimethacrylate, acting as crosslinking monomer
  • OEGDMA 550 01igo (ethylene glycol) dimethacrylate, acts as a crosslinking monomer
  • Biotin functioned as a recognition group immobilized in the porous photonic crystal, avidin as the analyte to be detected. At a molecular weight of about 66 kDa it was a biopolymer. Up to four biotin units bind selectively and with high binding constants (K - 10 15 ) to one avidin molecule. To detect avidin, biotin was covalently coupled to the porous photonic crystal. This was realized via a polymer-analogous esterification of the carboxyl group of the biotin with the hydroxyl group of the hydroxyethyl methacrylate.
  • DMAP Dimethylaminopyridine
  • EDC 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide
  • HOBt 1-hydroxybenzotriazole
  • NHS N-hydroxysuccinimide
  • FIG. 3 shows a hydrogel with biotin as biomolecule-specific recognition groups, wherein the hydrogel has a Bragg reflection due to its porous photonic crystal structure. Avidin is added, this binds to the
  • FIG. 4 shows the wavelengths of the color reflection of a responsive porous photonic crystal.
  • IHO-1 refers to the not yet biotinylated porous photonic crystal
  • bIHO-1 to the biotinylated porous photonic crystal before adding avidin
  • bIHO-1 + avidin to the biotinylated porous photonic crystal
  • the porous photonic hydrogel crystal was prepared under the abovementioned conditions according to Example 1.
  • the accessible biotin content of the film after Steglich esterification with DCC as coupling reagent was 0.01% relative to the hydroxyl groups present. that after addition of avidin to the biotinylated inverse opal (triangles) the peak wavelengths are red-shifted, the effect is most pronounced around room temperature, which is advantageous for a detection method of biomolecules.

Landscapes

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

Abstract

L'invention concerne un hydrogel sensible chimiquement réticulé, présentant une structure cristalline photonique poreuse et contenant des groupes reconnaissant spécifiquement les biomolécules.
EP14815644.1A 2014-01-08 2014-12-12 Hydrogel sensible pour mettre en évidence des biomolécules Withdrawn EP3092068A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE102014200135.8A DE102014200135A1 (de) 2014-01-08 2014-01-08 Responsives Hydrogel für den Nachweis von Biomolekülen
PCT/EP2014/077554 WO2015104139A1 (fr) 2014-01-08 2014-12-12 Hydrogel sensible pour mettre en évidence des biomolécules

Publications (1)

Publication Number Publication Date
EP3092068A1 true EP3092068A1 (fr) 2016-11-16

Family

ID=52134148

Family Applications (1)

Application Number Title Priority Date Filing Date
EP14815644.1A Withdrawn EP3092068A1 (fr) 2014-01-08 2014-12-12 Hydrogel sensible pour mettre en évidence des biomolécules

Country Status (4)

Country Link
US (1) US20170003281A1 (fr)
EP (1) EP3092068A1 (fr)
DE (1) DE102014200135A1 (fr)
WO (1) WO2015104139A1 (fr)

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107262079B (zh) * 2017-06-20 2019-08-27 湖南大学 一种用于同时监测和去除铀酰离子的智能光子晶体材料
DE102020200643A1 (de) * 2020-01-21 2021-07-22 Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung eingetragener Verein Durchlasskontrollelement sowie verfahren zu dessen betreiben
CN114149544B (zh) * 2021-12-16 2022-11-04 北京理工大学 一种粘性光子晶体水凝胶传感器及其制备方法及应用
CN114741875B (zh) * 2022-04-11 2024-06-18 西南石油大学 一种石英表面亲疏水基团定量修饰模型建立方法

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6753191B2 (en) 1996-11-06 2004-06-22 University Of Pittsburgh Polymerized crystalline colloidal array chemical sensing materials for use in high ionic strength solutions
WO2000000278A1 (fr) * 1998-06-26 2000-01-06 University Of Pittsburgh Of The Commonwealth System Of Higher Education Matieres d'hydrogel a reseau colloidal cristallin de poches d'eau permettant la detection et la separation de macromolecules
WO2008098339A1 (fr) * 2007-02-16 2008-08-21 The Governing Council Of The University Of Toronto Cristal photonique compressible
CA2753262A1 (fr) * 2009-02-25 2010-09-02 Opalux Incorporated Dispositif a cristal photonique reagissant a la temperature

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
None *
See also references of WO2015104139A1 *

Also Published As

Publication number Publication date
US20170003281A1 (en) 2017-01-05
DE102014200135A1 (de) 2015-07-09
WO2015104139A1 (fr) 2015-07-16

Similar Documents

Publication Publication Date Title
DE60121387T2 (de) Oberflächenbehandelter optischer Kunststoffgegenstand und Verfahren zur Oberflächenbehandlung
DE69531617T3 (de) Polymere mikrokugen und herstellungsverfahren
US20160002457A1 (en) Tunable and responsive photonic hydrogels comprising nanocrystalline cellulose
EP3092068A1 (fr) Hydrogel sensible pour mettre en évidence des biomolécules
DE3434822A1 (de) Membran fuer reagenztraegerschichten, verfahren zu deren herstellung, sowie deren verwendung in analytischen mitteln und analysenverfahren
CH665565A5 (de) Verfahren zur steuerung der konzentration einer eine makromolekulare verbindung enthaltenden waessrigen loesung oder emulsion.
DE102016105122B4 (de) Verfahren zur Synthese hybrider Kern-Schale-Mikropartikel umfassend einen Polymerkern und eine Siliziumdioxidschale mit kontrollierter Struktur und Oberfläche
CN110041464B (zh) 一种高强度光子晶体水凝胶及其制备方法和应用
DE102007024642A1 (de) Hydrogel-Implantat für Sensorik von Metaboliten am Auge
DE102004032430A1 (de) Verfahren zur Herstellung von molekular geprägten Polymeren
EP1721160B1 (fr) Procede pour immobiliser de maniere covalente des biomolecules sur des surfaces polymeriques
DE3040106C2 (de) Hydrophiles Füllmaterial für die Chromatographie
DE3407814A1 (de) Phasentraeger fuer die verteilungschromatographie von makromolekuelen, verfahren zu ihrer herstellung und ihre verwendung
EP3484612A1 (fr) Support chromatographique comprenant des microglobules liés et son procédé de fabrication
EP2191309B1 (fr) Cristaux photoniques à partir de particules polymères à interaction interparticulaire
DE112018000044B4 (de) Lichtdurchlässigkeitssteuerfolie
KR102025864B1 (ko) 아크릴산을 포함하는 하이드로젤 액추에이터 및 그 제조방법
Purkait et al. Temperature-responsive membranes
DE60033665T2 (de) Gemusterte Polymeroberflächen geeignet für Biokonjugationen und Verfahren zu ihrer Herstellung
DE3427441C2 (fr)
WO2021013814A1 (fr) Procédé pour étirer une molécule d'acide nucléique
Abbasi Moud Nanocellulose Product Design Aided by Confocal Laser Scanning Microscopy
EP1527203B1 (fr) Hydrogel réticulable par voie radicalaire comprenant des groupes de liaisons
DE102017117478A1 (de) Sensormaterial mit temperaturabhängiger Färbung
DE69920060T2 (de) Superabsorbierende materialien auf basis von butylgummi, verfahren zur herstellung und ihre verwendung

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 20160606

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR

AX Request for extension of the european patent

Extension state: BA ME

DAX Request for extension of the european patent (deleted)
STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: EXAMINATION IS IN PROGRESS

17Q First examination report despatched

Effective date: 20190509

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN

18D Application deemed to be withdrawn

Effective date: 20191120