US20060175193A1 - Polyelectrolyte complex(e.g.zwitterionic polythiophenes) with a receptor (e.g.polynucleotide, antibody etc.) for biosensor applications - Google Patents

Polyelectrolyte complex(e.g.zwitterionic polythiophenes) with a receptor (e.g.polynucleotide, antibody etc.) for biosensor applications Download PDF

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US20060175193A1
US20060175193A1 US10/514,191 US51419103A US2006175193A1 US 20060175193 A1 US20060175193 A1 US 20060175193A1 US 51419103 A US51419103 A US 51419103A US 2006175193 A1 US2006175193 A1 US 2006175193A1
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complex
polyelectrolyte
receptor
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Olle Inganas
Peter Asborg
Peter Nilsson
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    • 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
    • G01N33/566—Immunoassay; Biospecific binding assay; Materials therefor using specific carrier or receptor proteins as ligand binding reagents where possible specific carrier or receptor proteins are classified with their target compounds
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6813—Hybridisation assays
    • C12Q1/6816—Hybridisation assays characterised by the detection means
    • C12Q1/6825—Nucleic acid detection involving sensors
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6813—Hybridisation assays
    • C12Q1/6827—Hybridisation assays for detection of mutation or polymorphism
    • 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
    • G01N33/531—Production of immunochemical test materials
    • 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
    • G01N33/54366—Apparatus specially adapted for solid-phase testing

Definitions

  • the present invention relates to methods for detection of biomolecular interactions through the detection of alterations of the intra- and inter-chain processes in materials based on zwitterionic conjugated polyelectrolytes.
  • conjugated polymers such as poly(thiophene) and poly(pyrrole) can be used to couple analyte/receptor interactions, as well as non-specific interactions, into observable responses.
  • CPs based sensors are sensitive to very minor perturbations, due to amplification by a collective system response and therefore offer a key advantage compared to small molecules based sensors.
  • the possibility to use CPs as detecting elements for biological molecules requires that polymers are compatible with aqueous environment.
  • conjugated polyelectrolytes As recently used to detect biomolecules through their impact on the conditions for photoinduced charge or excitation transfer. Conjugated polyelectrolytes offer possibilities for very sensitive measurements, and may become ubiquitous for genomics and proteomics in the future, if the optical or electronic processes in these materials can be used to track biospecific interactions.
  • conjugated polymers can be modified by the introduction of suitable side chains in the 3-position.
  • Polythiophene derivatives that exhibit biotin and different carbohydrates has been synthesized and shown to undergo colorimetric transitions in response to binding of streptavidin and different types of bacteria and viruses, respectively.
  • the presently demonstrated systems use covalent attachment of a receptor to the side chains of the conjugated polymer. Therefore, methods without the need of covalent attachment of the receptor would be desirable, and such systems have been developed, see Boissinot, M., Leclerc, M, Ho, H-A. Patent Appl. WO02081735, 2002.
  • the object of the present invention is therefore to provide means and methods that meet these and other needs.
  • This object is in a first aspect achieved with a complex between a conjugated polyelectrolyte, and one or more receptor molecules specific for a target biomolecule analyte, said polyelectrolyte and said receptor being non-covalently bound to each other, usable as a probe for biomolecular interactions, defined in claim 1 .
  • the term “probe” shall be taken to mean any form of a complex as defined in claim 1 , capable of responding to biomolecular interactions occurring between a receptor in the complex and another species, such as molecules, cells, viruses, bacteria, spores, microorganisms, peptides, carbohydrates, nucleic acids, lipids, pharmaceuticals, antigens, antibodies, proteins, enzymes, toxins, any organic polymers or combination of these molecules that interacts with receptors of interest, by changing at least one property of the complex that can be detected by external means.
  • another species such as molecules, cells, viruses, bacteria, spores, microorganisms, peptides, carbohydrates, nucleic acids, lipids, pharmaceuticals, antigens, antibodies, proteins, enzymes, toxins, any organic polymers or combination of these molecules that interacts with receptors of interest, by changing at least one property of the complex that can be detected by external means.
  • the polyelectrolyte comprises copolymers or homopolymers of thiophene, pyrrole, aniline, furan, phenylene, vinylene or their substituted forms, and preferably the conjugated polyelectrolyte has one or more zwitterionic side chain functionalities.
  • a biosensor device for determining selected properties of biomolecules comprising a complex of the kind identified above, and a substrate for said complex in which said complex is exposable to said target analyte.
  • the biosensor device is defined in claim 14 .
  • a method of determining selected properties of biomolecules comprising exposing a complex as defined above, to a target biomolecule analyte whereby the analyte and the receptor interact, detecting a change of a property of said polyelectrolyte in response to the interaction between the receptor and the analyte; and using the detected change to determine said selected property of said biomolecule.
  • the method is defined in claim 17 .
  • the multiplicity of biomolecular interactions that one may wish to identify also implies that the invention in a still further aspect, can be implemented in the form of a microarray, and which calls for anchoring and patterning of the detecting system on a surface, defined in claim 22 .
  • FIG. 1 shows the chemical structure of poly(3-[(S)-5-amino-5-carboxyl-3-oxapentyl]-2,5-thiophenylene hydrochloride) (POWT), a zwitterionic polythiophene derivative.
  • FIG. 2 schematically illustrates the method according to the invention.
  • FIG. 3 shows the absorptionspectra of 1.16 ⁇ mol POWT (on a monomer basis) and 0 mol ( ⁇ ), 6.4 nmol ( ⁇ ) of an oligonucleotide (5′-CAT GAT TGA ACC ATC CAC CA-3′) after 5 minutes of incubation in 10 mM Na-phosphate buffer pH 7.5, or in the same buffer system with 6.4 nmol of a complementary oligonucleotide ( ⁇ ).
  • FIG. 4 shows the emission spectra of 23.1 nmol POWT (on a monomer basis) and 0 mol ( ⁇ ), 1.28 nmol ( ⁇ ), and 2.56 nmol (x) of an oligonucleotide (5′-CAT GAT TGA ACC ATC CAC CA-3′) after 5 minutes of incubation in 10 mM Na-phosphate buffer pH 7.5, or in the same buffer system with 1.28 nmol of a complementary oligonucleotide ( ⁇ ). All of the emission spectra were recorded with excitation at 400 nm.
  • FIG. 5 shows the emission spectra of 100 nmol POWT (on a monomer basis) (x) with 1.0 equivalent (on a monomer basis) of a positively charged peptide, JR2K ( ⁇ ), 1.0 equivalent of a negatively charged peptide, JR2E ( ⁇ ), 0.5 equivalents JR2E plus an addition of 0.5 equivalents JR2K ( ⁇ ), 0.5 equivalents JR2K plus an addition of 0.5 equivalents JR2E ( ⁇ ), 2.0 equivalent JR2K ( ⁇ ), and 2.0 equivalent JR2E ( ⁇ ) after 10 min of incubation in a 20 mM Na-phosphate buffer pH 7.4. All of the emission spectra were recorded with excitation at 400 nm.
  • FIG. 6 shows the Emission spectra of 26.1 nmol POWT in 20 mM Na-phosphate pH 7.5, upon addition of 4.9 ⁇ M of a synthetic peptide, with a receptor site for carbonic anhydrase (thin line), and after addition of 13 ⁇ M of carbonic anhydrase (bold line).
  • FIG. 7 shows the fluorescence images of POWT/DNA complexes. Hydrogels of POWT and single stranded DNA after binding of complementary DNA (bottom left) and non-complementary DNA (bottom right). Cross points (100 ⁇ 100 ⁇ m) of POWT and single stranded DNA after binding of complementary DNA (top left) or non-complementary DNA (top right). The fluorescence was recorded with an epifluorescence microscope (Zeiss Axiovert inverted microscope A200 Mot) equipped with a CCD camera (Axiocam HR).
  • FIG. 8 shows the DNA-hybridisation event on a POWT/gold chip monitored with a BiacoreX instrument Injection and wash out of (in order): ssDNA1 (characterization, 1540 RU), ssDNA1 (non-complementary, 30 RU), ssDNA2 (complementary, 860 RU). 0.15 M PBS buffer was used.
  • FIG. 9 shows the microcontact printing of POWT.
  • Table 1 shows the difference in ratio of emission intensity at the wavelengths 540 nm/585 nm and 540 nm/670 nm upon addition of 1.28 nmol of different oligonucleotides to a mixture of 23.1 nmol POWT and 1.28 nmol of a single stranded oligonucleotide.
  • Table 2 shows the absorption maximum and the ratio of the intensity of the emitted light at 540 nm/610 nm for POWT and POWT/peptide complexes after 10 min incubation in 20 mM Na-phosphate pH 7.4
  • the present invention relates to a novel complex between zwitterionic conjugated polyelectrolytes and a receptor, the polyelectrolyte acting as a carrier for said receptor, without the requirement to label the analytes or to covalently attach the receptors to the carrier.
  • the complex is used as a probe for responding to biomolecular interactions. It also relates to a biosensor device comprising such complex and a method for detection of molecular interactions.
  • the invention is based on zwitterionic polyelectrolyte forming a complex with one or more receptor molecules.
  • This complex is formed without covalent bonding and is based on hydrogen bonding, electrostatic- and non-polar interactions between the zwitterionic conjugated polymers and the receptor molecules, herein referred to as non-covalent bonding, which further includes any type of bonding that is not covalent in its nature.
  • the present invention utilizes changes of the zwitterionic conjugated polyelectrolyte/receptor molecules complex or alterations of the net charge of the receptor molecules, which induce conformational transitions of the backbone of the zwitterionic conjugated polyelectrolyte, separation or aggregation of zwitterionic conjugated polyelectrolyte chains. Furthermore, conformational transitions of the backbone of the zwitterionic conjugated polyelectrolyte, separation or aggregation of zwitterionic conjugad polyelectrolyte chains, alter the intra- and inter-chain processes of the zwitterionic conjugated polyelectrolytes. These changes can be detected in solution or on a surface.
  • the present invention allows, but is not limited to, detection of biospecific recognition through DNA (base pairing), proteins (antigen/antibody), glycoproteins or shorter purpose designed peptides.
  • the novel complex is suitably implemented as an active part of a biosensor device, e.g. by immobilizing the polyelectrolyte on a substrate in a biosensor cell.
  • the biosensor device comprises a suitable receptacle for said substrate, and a complex between polyelectrolyte and receptor is formed on the substrate.
  • the complex can be provided in solution and passed through a flow cell while an analyte solution is mixed with the flow of complex solution.
  • the interaction can be monitored by various analytical techniques.
  • poly(3-[(S)-5-amino-5-carboxyl-3-oxapentyl]-2,5-thiophenylene hydrochloride) (POWT, see FIG. 1 ) can be mentioned.
  • Studies of this polymer see Andersson, M.; Ekeblad, P. O.; Hjertberg, T.; Wennerström, O.; Inganäs, O. Polymer Commun.
  • the zwitterionic groups create versatile hydrogen bonding patterns with different molecules.
  • the present invention relates to a variety of conjugated polyelectrolytes, with a minimum of 5 mers, consisting of mers derived from the monomers thiophene, pyrrole, aniline, furan, phenylene, vinylene or their substituted forms, forming homopolymers and copolymers there from. Furthermore, monomers with anionic-, cationic or zwitterionic side chain functionalities are included within the scope of the invention.
  • the side chain functionalities is derived from, but not limited to, amino acids, amino acid derivatives, neurotransmittors, monosaccharides, nucleic acids, or combinations and chemically modified derivatives thereof.
  • the conjugated polyelectrolytes of the present invention may contain a single side chain functionality or may comprise two or more different side chain functionalities.
  • the zwitterionic groups create versatile hydrogen bonding patterns with different molecules.
  • the zwitterionic polyelectrolytes of the present invention form a complex with one or more receptor molecules ( FIG. 2 ).
  • This complex is formed without covalent bonding and based on hydrogen bonding, electrostatic- and non-polar interactions between the zwitterionic conjugated polymers and the receptor molecules.
  • the receptor molecules will act as the recognition site for analytes or as anchors for performing enzymatic reactions, such as phosphorylation.
  • a wide variety of receptor molecules can be used and the choice of molecule is only limited by the affinity to the conjugated polymers and the recognition properties of desirable analytes.
  • Appropriate receptor molecules include, but are not limited to, peptides, carbohydrates, nucleic acids, lipids, pharmaceuticals, antigens, antibodies, proteins, any organic polymers or combination of these molecules that are capable of interacting with analytes of interest.
  • the receptor molecules can be chemically modified to interact with the conjugated polymers of interest.
  • Methods of derivatizing a diverse range of compounds e.g. carbohydrates, proteins, nucleic acids and other chemical groups
  • amino acid side chains can easily be modified to contain polar and non-polar groups, or groups with hydrogen bonding abilities.
  • the conjugated polyelectrolyte/receptor molecules complex Upon binding of or exposure to one or more analytes, the conjugated polyelectrolyte/receptor molecules complex is subject to changes or alterations of the net charge of the receptor molecules.
  • Appropriate analytes include, but are not limited to, cells, viruses, bacteria, spores, microorganisms, peptides, carbohydrates, nucleic acids, lipids, pharmaceuticals, antigens, antibodies, proteins, enzymes, toxins, any organic polymers or combination of these molecules that interacts with receptors of interest.
  • the analytes can be chemically modified to interact with the receptor molecules of interest.
  • Methods of derivatizing a diverse range of compounds e. g. carbohydrates, proteins, nucleic acids and other chemical groups
  • amino acid side chains can easily be modified to contain polar and non-polar groups, or groups with hydrogen bonding abilities.
  • the conformational transitions of the backbone of the zwitterionic conjugated polyelectrolyte, separation or aggregation of polyelectrolyte chains will alter the intra- and inter-chain processes of the zwitterionic conjugated polyelectrolyte and can for example be detected as a change in the ratio of the intensities of the emitted light at two or more different wavelengths (see example 3).
  • the emission intensities can be recorded by a fluorometer and enhancement of the photon flow in the detector can increase the sensitivity. This can be achieved using a laser as the excitation source.
  • the fluorometric change can also be detected by the use of a fluorescence microscope or a confocal microscope.
  • a combination of excitation or emission filter can be used and the picture can be recorded by a CCD-camera (see example 7 and 9), video camera, regular camera or by a Polaroid camera.
  • the pictures can then be analyzed by image processing software on a computer, Image correlation spectroscopy (ICS) or by other means.
  • ICS Image correlation spectroscopy
  • the zwitterionic conjugated polymers can be immobilized inside a conducting polymer hydrogel matrix for example poly [3,4-(ethylenedioxy)thiophene]/poly(styrenesulfonicacid) (PEDOT/PSS). Changes in resistance, capacitance and inductance can then be tracked with the zwitterionic conjugated polyelectrolytes, receptor or analyte molecules in an aqueous environment.
  • a conducting polymer hydrogel matrix for example poly [3,4-(ethylenedioxy)thiophene]/poly(styrenesulfonicacid) (PEDOT/PSS). Changes in resistance, capacitance and inductance can then be tracked with the zwitterionic conjugated polyelectrolytes, receptor or analyte molecules in an aqueous environment.
  • SPR Surface plasmon resonance
  • Ellipsometry imaging or null ellipsometry
  • imaging or null ellipsometry is an optical technique that uses polarised light to sense the dielectric properties of a sample and can be used to detect these changes in thickness on a sub-angstrom level.
  • the interaction of receptor and analyte molecules with zwitterionic conjugated polyelectrolyte can also be detected by electrical and electrochemical methods.
  • a gel or network of the zwitterionic conjugated polyelectrolyte can be formed, and thus a three dimensional object is obtained where each polymer chain is in (indirect) contact with all chains in the network. If the zwitterionic conjugated polyelectrolyte is in a semiconducting state—such as when the luminescence properties is used—it will exhibit a rather low conductivity, which is somewhat difficult to easily distinguish from the ionic conductivity of the aqueous medium bathing the gel. It is therefore desirable to form highly conducting gels of the sensitive macromolecule that allow electrical conduction in the network.
  • a difficulty is that the doping of the conjugated chains, which gives a metallic polymer and a high conductivity, will not only turn on conductivity but also change the mechanical properties and geometry of the chains, thereby hindering the mechanism at work in the case of luminescence detection.
  • a solution to that problem is the use of two component polymer gels, where one component A gives the high conductivity and another component B the biospecific interactions. If these two compounds are combined in a suitable manner, the changes of geometry of the gel due to said interactions can be made to detect the interaction between component B and biomolecules.
  • Component A can be an aqueous dispersion of a highly doped polymer and component B, the zwitterionic conjugated polyelectrolyte can be combined, to make gels.
  • the intra- and inter-chain processes of the zwitterionic conjugated polyelectrolytes are altered by the interactions between receptor and analyte molecules or alteration of the net charge of the receptor molecule, and leads to changes of the electrochemical properties of the resulting complex, which can then be used to build electrochemical detectors for biomolecules.
  • a change of the redox potential of the hydrogel formed in the presence of a biomolecule can be used to detect the presence of a complementary biomolecule.
  • Similar devices, using conjugated polymers with a covalently attached receptor have been studied by Korri-Youssoufi, H., et al in “Toward bioelectronics: Specific DNA recognition based on an oligonucleotide-functionalized polypyrrole”, J. Am. Chem. Soc. 1997, 119, 7388-7389.
  • the zwitterionic conjugated polyelectrolytes, the zwitterionic conjugated polyelectrolyte/receptor molecules complex or the receptor molecules of the present invention can be immobilized on a variety of solid supports, including, but not limited to silicon wafers, glass (e.g. glass slides, glass beads, glass wafers etc.), silicon rubber, polystyrene, polyethylene, Teflon, silica gel beads, gold, indium tin oxide (ITO coated materials, e.g. glass or plastics), filter paper (e.g. nylon, cellulose and nitrocellulose), standard copy paper or variants and separation media or other chromatographic media.
  • solid supports including, but not limited to silicon wafers, glass (e.g. glass slides, glass beads, glass wafers etc.), silicon rubber, polystyrene, polyethylene, Teflon, silica gel beads, gold, indium tin oxide (ITO coated materials, e.g. glass or plastics), filter paper (e.
  • Transfer of the zwitterionic zwitterionic conjugated polyelectrolyte to the solid support can be achieved by using i.a. but not limited to, dip coating, spin-coating, contact printing, screen printing, ink jet technologies, spraying, dispensing and microfluidic printing by the use of soft lithography or the BiacoreTM (Biacore, Uppsala, Sweden) system.
  • Immobilization of the zwitterionic conjugated polyelectrolytes is achieved by physical adhesion to the solid support at elevated temperatures or by entrapment in a hydrogel matrix.
  • Immobilization of the zwitterionic conjugated polyelectrolytes of the present invention may be desired to improve their ease of use, assembly into devices (e.g. arrays or parallel lines), stability, robustness, fluorescent response, to fit into the process of high-throughput-screening (HTS) using micro titer plates and other desired properties.
  • HTS high-throughput-screening
  • the receptor molecules of the present invention can be immobilized together with the zwitterionic conjugated polyelectrolyte (i.e. mixed with the polyelectrolyte solution). Another way to immobilize the receptor molecules is to place them underneath or on top of the zwitterionic conjugated polyelectrolyte. Transfer of the receptor molecules mixed together with zwitterionic conjugated polyelectrolyte to the solid support can be achieved by, but not limited to, using dip coating, spin-coating, contact printing, screen printing, ink jet technologies, spraying, dispensing and microfluidic printing (see example 9) by the use of soft lithography (see example 10) or the Biacore TM system (see example 8).
  • the receptor molecules If the receptor molecules is to be placed underneath the zwitterionic zwitterionic conjugated polyelectrolyte it has to be transferred to the solid support in the same way as it would have been mixed together with the polyelectrolyte as mentioned above. Placing the receptor molecules on top of the zwitterionic conjugated polyelectrolyte is done in the same way but after the polyelectrolyte has been immobilized to the solid support.
  • the receptor molecules will act as the recognition site for analytes or as anchors for performing enzymatic reactions, such as phosphorylation.
  • Solvents for the zwitterionic conjugated polyelectrolytes of the present invention and the receptor molecules during the immobilization to the solid support can be, but are not limited to, water, buffered water solutions, methanol, ethanol and combinations thereof. Supporting polymers of other kinds can also be added in this step.
  • the receptor molecules When the receptor molecules are immobilized on the solid support underneath, on top of or together with the zwitterionic conjugated polyelectrolyte of the present invention they form a complex with the polyelectrolyte through non-covalent interactions ( FIG. 2 ). This complex is formed without covalent chemistry and is based on hydrogen bonding, electrostatic- and non-polar interactions between the zwitterionic conjugated polyelectrolyte and the receptor molecule. Immobilization of the receptors to the zwitterionic conjugated polyelectrolytes of the present invention may be desired to improve their ease of use, assembly into devices (e.g.
  • receptor molecules have been immobilized onto cationic or anionic conjugated polymers for detection of analytes [15], prior to the the present invention, immobilization without covalent chemistry and based on hydrogen bonding, electrostatic- and non-polar interactions between the zwitterionic conjugated polyelectrolytes and the receptor molecules had not been realized.
  • the zwitterionic conjugated polyelectrolyte and receptor molecules can be entrapped inside polymer matrices on top of a solid support or free floating in solution.
  • a gel or network of the zwitterionic conjugated polymers can be formed, where each zwitterionic conjugated polyelectrolyte chain of the present invention is in (indirect) contact with all chains in the network.
  • Realization of these polymer matrices can be done by mixing c zwitterionic conjugated polyelectrolyte with other polymers such as, but not limited to, poly [3,4-(ethylenedioxy) thiophene]/poly(styrenesulfonicacid) (PEDOT/PSS), poly (diallyldimethylammonium chloride) (PDADMAC), poly-4-vinylpyridine (PVPy), poly(pyrrole) (PPy), poly(vinylalcohol) (PVA), poly(aniline) (PANI) or combinations thereof.
  • PDAMAC poly (diallyldimethylammonium chloride)
  • PVPy poly-4-vinylpyridine
  • PVPy poly(pyrrole)
  • PVA poly(vinylalcohol)
  • PANI poly(aniline)
  • the zwitterionic conjugated polyelectrolytes of the present invention can be mixed together with these polymers before immobilization to the solid support or transferred afterwards.
  • Receptor molecules of interest can be transferred together with the zwitterionic conjugated polyelectrolyte or in a subsequent step.
  • a microarray or parallel line format can be used if desired, necessary or for other reasons.
  • this network or hydrogel approach can be used to detect conformational changes and aggregation of the zwitterionic conjugated polyelectrolyte chains due to interaction between receptor and analyte molecules or change in the net charge of the receptor molecules. These alterations can then be detected by measuring absorption, fluorescence, electrical properties, impedance or by other means.
  • the generation of large arrays or parallel lines of the zwitterionic conjugated polyelectrolytes with the same or different receptor molecules in each spot or line can overcome shortcomings of a single sensor or a solution based approach.
  • the array or parallel line approach opens up the parallel analysis of one or different analytes to one or different receptors in an easy way.
  • the main purpose of using arrays or lines is to increase ease of use, portability, quantification, selectivity among other qualities and characteristics. With this approach we can explore the ability to measure multicomponent samples and to use partially selective sensor spots. This gives the opportunity to analyse two or more samples of interest at the same time, to do on-chip concentration determinations and to study the background.
  • immobilizing the zwitterionic conjugated polyelectrolyte and/or the receptor molecules on solid supports of any size and in any chosen patterns (such as arrays, lines, spots, posts) small, portable, easily read and interpretable devices can be constructed.
  • each individual detector by the simple blending of the zwitterionic conjugated polyelectrolyte and biomolecules, we have removed the necessity of covalent chemistry for making each one of many thousands of detectors in a detector array (microarray).
  • the zwitterionic conjugated polyelectrolyte and zwitterionic conjugated polyelectrolyte/biomolecule complexes can be printed by micro contact printing using elastomer stamps ( FIG.
  • Transfer onto a microarray surface may also be done by spotting zwitterionic conjugated polyelectrolyte solutions, or by ink jetting polyelectrolyte solutions or by the other methods mentioned above. These steps are essential to prepare a multipixel microarray.
  • a stock solution containing 0.5 mg ml ⁇ 1 POWT in de-ionised water was prepared and incubated for 30 minutes. 50 ⁇ l of the polymer solution was mixed with 64 ⁇ l of DNA-solution (100 nmol ml ⁇ 1 , 5′-CAT GAT TGA ACC ATC CAC CA-3′, purchased from SGSDNA, Köping, Sweden).
  • a stock solution containing 0.5 mg ml ⁇ 1 POWT in de-ionised water was prepared and incubated for 30 minutes. 10 ⁇ l of the polymer solution was mixed with 12.8 ⁇ l of DNA-solution (100 nmol ml ⁇ 1 , 5′-CAT GAT TGA ACC ATC CAC CA-3′, purchased from SGSDNA, Köping, Sweden).
  • a stock solution containing 0.5 mg ml ⁇ 1 POWT in de-ionised water was prepared and incubated for 30 minutes. 10 ⁇ l of the polymer solution was mixed with 12.8 ⁇ l of DNA-solution (100 nmol ml ⁇ 1 , 5′-CAT GAT TGA ACC ATC CAC CA-3′, purchased from SGSDNA, Köping, Sweden).
  • the samples were diluted with de-ionised water, a stock buffer solution (Na-phosphate pH 7.5 and a 1.0 equivalent amount of the respective nucleotide (5′-TGG TGG ATG GTT CAA TCA TG-3′, 5′-TGG TGG ATG CTT CAA TCA TG -3′, 5′-TGG TGG AAC GTT CAA TCA TG-3′, 5′-TGG TGG AAC CTT CAA TCA TG -3′ or 5′-CAT GAT TGA ACC ATC CAC CA -3′, purchased from SGSDNA, Köping, Sweden) to a final volume of 1500 ⁇ l containing 10 mM Na-phosphate.
  • a stock buffer solution Na-phosphate pH 7.5 and a 1.0 equivalent amount of the respective nucleotide
  • the samples were incubated for 5 minutes and the emission spectra were recorded with a ISA Jobin-Yvon spex FluoroMax-2 apparatus.
  • the difference in ratio of emission intensity at the wavelengths 540 nm/585 nm and 540 nm/670 nm were calculated.
  • the emitted light at 540 nm and 585 nm is due to intra-chain processes and the emitted light at 670 nm is due to an inter-chain process (aggregation of POWT chains).
  • Nucleotides with one, two or three mismatches can easily be detected, as the difference in ratio of the emission intensity at the wavelengths 540 nm/585 nm and 540 nm/670 nm are influenced by the degree of mismatch between the DNA strands (Table 1).
  • a stock solution containing 3.7 mg ml ⁇ 1 POWT in de-ionised water was prepared and incubated for 30 minutes. 10 ⁇ l of the polymer solution was mixed with 10 ⁇ l or 20 ⁇ l of a negatively charged peptide (NH2-N-A-A-D-L-E-K-A-l-E-A-L-E-K-H-L-E-A-K-G-P-V-D-A-A-Q-L-E-K-Q-L-E-Q-A-F-E-A-F-E-R-A-G-COOH) or the positively charged peptide (NH2-N-A-A-D-L-K-K-I-K-A-L-K-K-H-L-K-A-K-G-P-V-D-A-A-Q-L-K-K-Q-L-K-Q-A-F-K-A-F-K-R-A-G-COOH) solution (2.2 mg
  • the samples were diluted with a stock buffer solution (Na-phosphate pH 7.4) and 10 ⁇ l de-ionised water or 10 ⁇ l of the positive/negative peptide solution (2.2 mg ml ⁇ 1 ) to a final volume of 2000 ⁇ l containing 20 mM Na-phosphate.
  • a stock buffer solution Na-phosphate pH 7.4
  • 10 ⁇ l de-ionised water or 10 ⁇ l of the positive/negative peptide solution (2.2 mg ml ⁇ 1 ) to a final volume of 2000 ⁇ l containing 20 mM Na-phosphate.
  • a stock solution containing 3.7 mg ml ⁇ 1 POWT in de-ionised water was prepared and incubated for 30 minutes.
  • 10 ⁇ l of the polymer solution was mixed with 10 ⁇ l or 20 ⁇ l of a negatively charged peptide (NH2-N-A-A-D-L-E-K-A-I-E-A-L-E-K-H-L-E-A-K-G-P-V-D-A-A-Q-L-E-K-Q-L-E-Q-A-F-E-A-F-E-R-A-G-COOH) or a positively charged peptide (NH2-N-A-A-D-L-K-K-I-K-A-L-K-K-H-L-K-A-K-G-P-V-D-A-A-Q-L-K-K-Q-L-K-Q-A-F-K-A-F-K-R-A-G-COOH) solution (2.2
  • the samples were diluted with a stock buffer solution (Na-phosphate pH 7.4) and 10 ⁇ l de-ionised water or 10 ⁇ l of the positive/negative peptide solution (2.2 mg ml ⁇ 1 ) to a final volume of 2000 ⁇ l containing 20 mM Na-phosphate.
  • the samples were incubated for 10 minutes in room temperature and the emission spectra ( FIG. 5 , Table 2) were recorded with an ISA Jobin-Yvon spex FluoroMax-2 apparatus.
  • JR2K will shift the emission maximum to shorter wavelengths and increase the intensity of the emitted light, indicative of a non-planar POWT backbone and separation of POWT chains
  • addition of JR2E will shift the emission maximum to longer wavelengths and decrease the intensity of emitted light, indicative of a planar POWT backbone and aggregation of POWT chain
  • JR2E and JR2K has been tailor made to form a four-helix bundle and the formation of this structure can be detected by a change of the emission maximum and the intensity of the emitted light from POWT ( FIG. 5 ).
  • the emitted light at 540 nm and 610 nm is due to intra-chain processes and the emitted light at 670 nm is due to an inter-chain process (aggregation of polymer chains).
  • the difference in ratio of the emission intensity at the wavelengths 540 nm/610 nm and 540 nm/670 nm are influenced, as the different complexes between POWT and the different peptides are formed (Table 2).
  • the intra- and inter-chain processes of the POWT/JR2E (receptor) are clearly altered upon addition of JR2K (analyte).
  • a stock solution containing 0.5 mg ml ⁇ 1 POWT in de-ionised water was prepared and incubated for 30 minutes. 10 ⁇ l of the polymer solution was mixed with 5 ⁇ l of a negatively charged peptide solution (NH2-N-A-A-D-L-E-K-A-I-E-A-L-E-K-H-L-E-A-K-G-P-V-D-A-A-Q-L-E-K-Q-L-E-Q-A-F-E-A-F-E-R-A-G-COOH, modified with a receptor for carbonic anhydrase) (2.2 mg ml ⁇ 1 ), and diluted with de-ionised water to a final volume of 100 ⁇ l.
  • a negatively charged peptide solution (NH2-N-A-A-D-L-E-K-A-I-E-A-L-E-K-H-L-E-A-K-G-P-V-D-
  • 0.5 ⁇ l droplets of POWT (0.5 mg ml ⁇ 1 ) were placed on a polystyrene surface and left to dry for 10 min.
  • the polymer droplets were cross-linked with 0.5 ⁇ l DNA solution containing 0.5 equivalents on a monomer basis of 5′-AGA TTG GCG CAT TAC GAG GTT AGA-3′ or 5′-TCT AAC CTC GTA ATG CGC CAA TCT-3′ (purchased from SGSDNA, Köping, Sweden), respectively.
  • the fluorescence from the spots was recorded with an epifluorescence microscope (Zeiss Axiovert inverted microscope A200 Mot) equipped with a CCD camera (Axiocam HR), using a 405/30 nm bandpass filter (LP450, exposure time: 1500 ms), a 470/40 nm bandpass filter (LP515, exposure time: 1500 ms) and a 546/12 nm bandpass filter (LP590, exposure time: 500 ms).
  • LP450 405/30 nm bandpass filter
  • LP590 exposure time: 500 ms
  • a bare gold sensor chip was spin casted (1000 rpm, 30 s) with a 5 mg/ml solution of POWT in milliQ water. The film were annealed by heating the chip at 75° C. for 5 min. Finally, the chip was assembled on the sensor chip support by using glue or adhesive strips. Generally an injection sequence consisting of three injections were performed.
  • the first injection aims to characterize the polymer with ssDNA(5′-AGA TTG GCG CAT TAC GAG GTT AGA-3′, purchased from SGSDNA, Köping, Sweden), the second to verify that no unspecific binding occurs and the final injection aims to prove specific binding in the form of DNA hybridisation using 5′-AGA TTG GCG CAT TAC GAG GTT AGA-3′ or 5′-TCT AAC CTC GTA ATG CGC CAA TCT-3′ (purchased from SGSDNA, Köping, Sweden), respectively.
  • the polymer films were first swollen in degassed milliQ water and then equilibrated in degassed 20 mM phosphate pH 7,4 buffer (PBS) with salt concentrations (NaCl) ranging from 0 to 1 M.
  • PBS phosphate pH 7,4 buffer
  • NaCl salt concentrations
  • the injected DNA was solved in the same buffer as the running buffer and the concentration was usually around 1 ⁇ M.
  • the temperature was set to 25° C. during all experiments.
  • the hybridisation event was monitored with a BiacoreX instrument from Biacore AB (Uppsala, Sweden). The instrument has two flow channels with the approximate size of 0.5 ⁇ 2.5 mm. Manual loading is required and the maximal injection volume is 100 ⁇ l. As shown in FIG.
  • Sylgard 184 (Dow Corning, UK), a two component silicone rubber (poly(dimethylsiloxane), PDMS), was used for preparing elastomer stamps used for transferring POWT to solid surfaces.
  • the prepolymer and the curing agent is mixed according to the instructions provided by the manufacturer. This is then poured on templates prepared by photolithography using the negative photoresist SU-8 (Micro Chem Inc., Newton, Mass., USA) as the structural element on top of silicon wafers. Curing is accomplished by heating to 130° C. for at least 20 min.
  • the height of structures was 18 micrometer, and the substrate was a Si wafer cleaned in a boiling aqueous solution containing 5% each of ammonia and H 2 O 2 (TL-1 wash).
  • the geometry for templates was designed in CleWin Version 2.51 (WieWeb Software), and transferred to a Cr mask, which was used in the photolithography step.
  • silanization (dimethyl-dicholorosilane) was done to obtain the proper surface energy of the SU-8 template.
  • a solution of POWT (10 mg ml ⁇ 1 in methanol) was spin coated (2600 rpm) on to a glass surface previously cleaned by a TL-1 wash and modified by a 10 sec oxygen plasma treatment.
  • a PDMS stamp with 100 ⁇ m wide channels was modified by 10 sec oxygen plasma treatment and then placed onto the polymer film.
  • the channels were filled with the desired nucleotide solution (20 nmol 5′-AGA TTG GCG CAT TAC GAG GTT AGA-3 ′ or 5′-TCT AAC CTC GTA ATG CGC CAA TCT -3′ in deionised water) and then left to dry in room temperature before the stamp was removed.
  • Sylgard 184 (Dow Corning, UK), a two component silicone rubber (poly(dimethylsiloxane), PDMS), was used for preparing elastomer stamps used for transferring POWT to solid surfaces.
  • the prepolymer and the curing agent is mixed according to the instructions provided by the manufacturer. This is then poured on templates prepared by photolithography using the negative photoresist SU-8 (Micro Chem Inc., Newton, Mass., USA) as the structural element on top of silicon wafers. Curing is accomplished by heating to 130° C. for at least 20 min.
  • the height of structures was 18 micrometer, and the substrate was a Si wafer cleaned in a boiling aqueous solution containing 5% each of ammonia and H 2 O 2 (TL-1 wash).
  • the geometry for templates was designed in CleWin Version 2.51 (WieWeb Software), and transferred to a Cr mask, which was used in the photolithography step.
  • silanization (dimethyl-dicholorosilane) was done to obtain the proper surface energy of the SU-8 template.
  • the PDMS stamps were plasma treated for ⁇ 10 sec before being dip-coated in a water-based solution of POWT (5 mg ml ⁇ 1 ). The polymer was dried on the top of the stamp with N 2 .
  • the stamp was put face down for 20-25 minutes, on a glass substrate previously cleaned with a TL-1 wash, or a polystyrene surface modified by a 10 sec oxygen plasma treatment. Both substrates were moistened before stamp contact. After removal of the stamp, POWT had partly transferred to the glass as shown in FIG. 9 .
  • the detection capability of recognizing another DNA molecule is utilized.
  • the conjugated zwitterionic polymer is mixed together with a 0.1 equivalent amount (on a monomer basis) of a single stranded oligonucleotide in deionized water.
  • Gold electrodes which can be patterned in any way if desired, on a glass support is cleaned with ethanol. On top of these electrodes is a dispersion of a conducting polymer (PEDOT-PSS, commercial name Baytron from Bayer AG) is deposited.
  • the zwitterionic polymer/oligonucleotide complex is transferred on to the polymer surface by solution casting, contact printing, ink-jet printing or in other ways.
  • the resulting layer is analyzed using 2- or 4-point resistance measurement, by electrochemical methods or by impedance spectroscopy.

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Cited By (23)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20060204984A1 (en) * 2005-01-31 2006-09-14 The Regents Of University Of California Methods and compositions for aggregant detection
US20060216734A1 (en) * 2005-01-10 2006-09-28 The Regents Of The University Of California Methods and articles for strand-specific polynucleotide detection with cationic multichromophores
US20070088130A1 (en) * 2003-09-17 2007-04-19 The Regents Of The University Of California Conformationally Flexible Cationic Conjugated Polymers
US20070196819A1 (en) * 2004-03-24 2007-08-23 Biochromix Ab Patterning Method For Biosensor Applications And Devices Comprising Such Patterns
US20080064042A1 (en) * 2002-08-26 2008-03-13 Bazan Guillermo C Compositions for detection and analysis of polynucleotides using light harvesting multichromophores
US20080293164A1 (en) * 2006-10-06 2008-11-27 Sirigen Inc. Fluorescent Methods and Materials for Directed Biomarker Signal Amplification
WO2008143731A2 (en) * 2007-03-01 2008-11-27 University Of Florida Research Foundation, Inc. Surface grafted conjugated polymers
US20100112719A1 (en) * 2008-11-06 2010-05-06 Amihood Doron Electronic signal amplification in field effect device based chemical sensors
US7781172B2 (en) * 2003-11-21 2010-08-24 Kimberly-Clark Worldwide, Inc. Method for extending the dynamic detection range of assay devices
KR100997411B1 (ko) 2008-04-17 2010-11-30 부산대학교 산학협력단 반대이온의 교환을 이용한 공액 고분자 ｆｒｅｔ 시스템 및바이오센서
US8362193B2 (en) 2010-01-19 2013-01-29 Sirigen Group Limited Reagents for directed biomarker signal amplification
US8969509B2 (en) 2009-06-26 2015-03-03 Sirigen, Inc. Signal amplified biological detection with conjugated polymers
WO2015054484A1 (en) * 2013-10-09 2015-04-16 The University Of Akron Integrated zwitterionic conjugated polymers for bioelectronics, biosensing, regenerative medicine, and energy applications
US9371559B2 (en) 2002-06-20 2016-06-21 The Regents Of The University Of California Compositions for detection and analysis of polynucleotides using light harvesting multichromophores
US9527806B2 (en) 2010-07-13 2016-12-27 Stc.Unm Structure, synthesis, and applications for poly (phenylene) ethynylenes (PPEs)
US9549549B2 (en) 2011-08-03 2017-01-24 Stc.Unm Antimicrobial materials and methods
US9750250B2 (en) 2015-01-14 2017-09-05 Stc.Unm Conjugated polyelectrolytes and methods of using the same
US9968698B2 (en) 2013-11-08 2018-05-15 Stc. Unm Charged singlet-oxygen sensitizers and oppositely-charged surfactants
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US10307513B2 (en) * 2015-06-12 2019-06-04 University of Pittsburgh—of the Commonwealth System of Higher Education Biomimetic hydrogel scaffolds and related methods
US10533991B2 (en) 2014-03-14 2020-01-14 Stc.Unm P-phenylene ethynylene compounds as bioactive and detection agents
US10772851B2 (en) 2017-02-03 2020-09-15 Aaron Kurt Neumann Treatment and prevention of fungal infections
WO2024161291A1 (en) * 2023-01-30 2024-08-08 Asima Health Inc. Impedance-based liquid biopsy system and method for detecting and screening cancer

Families Citing this family (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
SE0401219D0 (sv) * 2004-05-10 2004-05-10 Biochromix Ab Metoder för detektera konformationsförändringar eller aggregering hos proteiner med hjälp av konjugerade polyelektrolyter
SE0401649D0 (sv) * 2004-06-28 2004-06-28 Olle Werner Inganaes Metoder för att konstruera elektroniska komponenter baserade på biomolekyler och konjugerande polymerer
EP1931979B1 (de) 2005-10-04 2015-11-18 Headway Technologies, Inc. Mikrofluidischer nachweis von analyten
US7878644B2 (en) 2005-11-16 2011-02-01 Gerber Scientific International, Inc. Light cure of cationic ink on acidic substrates
JP4510766B2 (ja) * 2006-01-31 2010-07-28 日本電信電話株式会社 細胞外電極
JP2010530955A (ja) * 2006-12-29 2010-09-16 ユニヴァーシティ オブ ワシントン 二重機能性非汚染表面および材料
WO2008130296A1 (en) 2007-04-18 2008-10-30 Biochromix Ab Binding of pathological forms of proteins using conjugated polyelectrolytes
KR101574251B1 (ko) * 2014-12-17 2015-12-03 주식회사 피엔에스테크놀로지 생화학물질 고정용 유기/무기 나노 복합체, 이의 제조방법 및 이를 포함하는 바이오 센서 또는 흡착 장치
EP4183853B1 (de) 2016-04-15 2024-10-02 Beckman Coulter, Inc. Photoaktive makromoleküle und ihre verwendung
US11787952B2 (en) 2017-05-17 2023-10-17 Abf Technologies, Llc Zwitterionic monomers, polyzwitterionic polymers formed therefrom, surface functionalization and surface modification
KR102347808B1 (ko) * 2020-12-03 2022-01-07 연세대학교 산학협력단 공액 고분자 기반 바이러스 검출용 복합체

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5670381A (en) * 1988-01-29 1997-09-23 Abbott Laboratories Devices for performing ion-capture binding assays
US6096825A (en) * 1994-04-22 2000-08-01 Bio Merieux Electrically conductive electroactive functionalized conjugated polymers, and uses thereof
US20010026915A1 (en) * 1992-11-13 2001-10-04 The Regents Of The University Of California Colorimetric glycopolythiophene biosensors
US6326936B1 (en) * 1997-07-22 2001-12-04 Thin Film Electronics Asa Electrode means, comprising polymer materials, with or without functional elements and an electrode device formed of said means
US20030124739A1 (en) * 2001-12-24 2003-07-03 Kimberly-Clark Worldwide, Inc. Polyelectrolytic internal calibration system of a flow-through assay
US6737279B1 (en) * 2001-03-28 2004-05-18 The Regents Of The University Of California Tuning the properties of conjugated polyelectrolytes and application in a biosensor platform

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH10505904A (ja) * 1994-09-13 1998-06-09 バイオサーキッツ コーポレイション 被検体を測定することにおける脂質層における分光光度的変化の直接的及び間接的変調
WO1996025665A1 (en) * 1995-02-13 1996-08-22 The Regents Of The University Of California Three-dimensional colorimetric assay assemblies
WO1999067423A1 (en) * 1998-06-22 1999-12-29 The Regents Of The University Of California Nucleic acid-coupled colorimetric analyte detectors
CA2330937A1 (en) * 1998-06-22 1999-12-29 Regents Of The University Of California Nucleic acid-coupled colorimetric analyte detectors
WO2000031750A1 (en) * 1998-11-19 2000-06-02 Bio Merieux Electrically conductive electroactive functionalized conjugated polymers, and uses thereof
JP2004528007A (ja) * 2000-09-29 2004-09-16 インサイト・ゲノミックス・インコーポレイテッド Ｇタンパク質共役受容体
EP1373246B1 (de) * 2001-04-05 2012-09-12 Geneohm Sciences Canada, Inc. Erkennung von negativ-geladenen polymeren mit hilfe von wasserlöslichen kationischen polythiophenderivaten

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5670381A (en) * 1988-01-29 1997-09-23 Abbott Laboratories Devices for performing ion-capture binding assays
US20010026915A1 (en) * 1992-11-13 2001-10-04 The Regents Of The University Of California Colorimetric glycopolythiophene biosensors
US6096825A (en) * 1994-04-22 2000-08-01 Bio Merieux Electrically conductive electroactive functionalized conjugated polymers, and uses thereof
US6326936B1 (en) * 1997-07-22 2001-12-04 Thin Film Electronics Asa Electrode means, comprising polymer materials, with or without functional elements and an electrode device formed of said means
US6737279B1 (en) * 2001-03-28 2004-05-18 The Regents Of The University Of California Tuning the properties of conjugated polyelectrolytes and application in a biosensor platform
US20030124739A1 (en) * 2001-12-24 2003-07-03 Kimberly-Clark Worldwide, Inc. Polyelectrolytic internal calibration system of a flow-through assay

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US10001475B2 (en) 2002-06-20 2018-06-19 The Regents Of The University Of California Light harvesting multichromophore compositions and methods of using the same
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USRE47874E1 (en) 2002-08-26 2020-02-25 The Regents Of The University Of California Methods and compositions for detection and analysis of polynucleotides using light harvesting multichromophores
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US9085799B2 (en) 2002-08-26 2015-07-21 The Regents Of The University Of California Methods and compositions for detection and analysis of polynucleotides using light harvesting multichromophores
US8841072B2 (en) 2002-08-26 2014-09-23 The Regents Of The University Of California Methods and compositions for detection and analysis of polynucleotides using light harvesting multichromophores
US20080064042A1 (en) * 2002-08-26 2008-03-13 Bazan Guillermo C Compositions for detection and analysis of polynucleotides using light harvesting multichromophores
USRE46817E1 (en) 2002-08-26 2018-05-01 The Regents Of The University Of California Methods and compositions for detection and analysis of polynucleotides using light harvesting multichromophores
US7914984B2 (en) 2003-09-17 2011-03-29 The Regents Of The University Of California Conformationally flexible cationic conjugated polymers
US20070088130A1 (en) * 2003-09-17 2007-04-19 The Regents Of The University Of California Conformationally Flexible Cationic Conjugated Polymers
US7781172B2 (en) * 2003-11-21 2010-08-24 Kimberly-Clark Worldwide, Inc. Method for extending the dynamic detection range of assay devices
US20070196819A1 (en) * 2004-03-24 2007-08-23 Biochromix Ab Patterning Method For Biosensor Applications And Devices Comprising Such Patterns
US7811755B2 (en) 2005-01-10 2010-10-12 The Regents Of The University Of California Methods and articles for strand-specific polynucleotide detection with cationic multichromophores
US20060216734A1 (en) * 2005-01-10 2006-09-28 The Regents Of The University Of California Methods and articles for strand-specific polynucleotide detection with cationic multichromophores
US20060204984A1 (en) * 2005-01-31 2006-09-14 The Regents Of University Of California Methods and compositions for aggregant detection
US7666594B2 (en) 2005-01-31 2010-02-23 The Regents Of The University Of California Methods for assaying a sample for an aggregant
US8354239B2 (en) 2006-10-06 2013-01-15 Sirigen, Inc. Fluorescent methods and materials for directed biomarker signal amplification
US8802450B2 (en) 2006-10-06 2014-08-12 Sirigen, Inc. Fluorescent methods and materials for directed biomarker signal amplification
US10641777B2 (en) 2006-10-06 2020-05-05 Sirigen, Inc. Fluorescent methods and materials for directed biomarker signal amplification
US20080293164A1 (en) * 2006-10-06 2008-11-27 Sirigen Inc. Fluorescent Methods and Materials for Directed Biomarker Signal Amplification
US10126302B2 (en) 2006-10-06 2018-11-13 Sirigen Ii Limited Fluorescent methods and materials for directed biomarker signal amplification
US11209438B2 (en) 2006-10-06 2021-12-28 Becton, Dickinson And Company Fluorescent methods and materials for directed biomarker signal amplification
US8158444B2 (en) 2006-10-06 2012-04-17 Sirigen, Inc. Fluorescent methods and materials for directed biomarker signal amplification
US9383353B2 (en) 2006-10-06 2016-07-05 Sirigen Inc. Fluorescent methods and materials for directed biomarker signal amplification
US11639937B2 (en) 2006-10-06 2023-05-02 Sirigen Ii Limited Fluorescent methods and materials for directed biomarker signal amplification
US10107818B2 (en) 2006-10-06 2018-10-23 Sirigen Ii Limited Fluorescent methods and materials for directed biomarker signal amplification
US10859578B2 (en) 2006-10-06 2020-12-08 Skigen, Inc. Fluorescent methods and materials for directed biomarker signal amplification
US8455265B2 (en) 2007-03-01 2013-06-04 Stc.Unm Surface grafted conjugated polymers
US20110159605A1 (en) * 2007-03-01 2011-06-30 University Of Florida Research Foundation, Inc. Surface Grafted Conjugated Polymers
WO2008143731A3 (en) * 2007-03-01 2009-04-30 Univ Florida Surface grafted conjugated polymers
WO2008143731A2 (en) * 2007-03-01 2008-11-27 University Of Florida Research Foundation, Inc. Surface grafted conjugated polymers
KR100997411B1 (ko) 2008-04-17 2010-11-30 부산대학교 산학협력단 반대이온의 교환을 이용한 공액 고분자 ｆｒｅｔ 시스템 및바이오센서
US20100112719A1 (en) * 2008-11-06 2010-05-06 Amihood Doron Electronic signal amplification in field effect device based chemical sensors
US8969509B2 (en) 2009-06-26 2015-03-03 Sirigen, Inc. Signal amplified biological detection with conjugated polymers
US8362193B2 (en) 2010-01-19 2013-01-29 Sirigen Group Limited Reagents for directed biomarker signal amplification
US10641775B2 (en) 2010-01-19 2020-05-05 Sirigen Ii Limited Reagents for directed biomarker signal amplification
US10094838B2 (en) 2010-01-19 2018-10-09 Sirigen Ii Limited Reagents for directed biomarker signal amplification
US12111321B2 (en) 2010-01-19 2024-10-08 Sirigen Ii Limited Reagents for directed biomarker signal amplification
US11899018B2 (en) 2010-01-19 2024-02-13 Sirigen Ii Limited Reagents for directed biomarker signal amplification
US10288620B2 (en) 2010-01-19 2019-05-14 Sirigen Ii Limited Reagents for directed biomarker signal amplification
US10302648B2 (en) 2010-01-19 2019-05-28 Sirigen Ii Limited Reagents for directed biomarker signal amplification
US11874278B2 (en) 2010-01-19 2024-01-16 Sirigen Ii Limited Reagents for directed biomarker signal amplification
US8455613B2 (en) 2010-01-19 2013-06-04 Sirigen Group Limited Reagents for directed biomarker signal amplification
US10365285B2 (en) 2010-01-19 2019-07-30 Sirigen Ii Limited Reagents for directed biomarker signal amplification
US8575303B2 (en) 2010-01-19 2013-11-05 Sirigen Group Limited Reagents for directed biomarker signal amplification
US10458989B2 (en) 2010-01-19 2019-10-29 Sirigen Ii Limited Reagents for directed biomarker signal amplification
US10481161B2 (en) 2010-01-19 2019-11-19 Sirigen Ii Limited Reagents for directed biomarker signal amplification
US11333666B2 (en) 2010-01-19 2022-05-17 Sirigen Ii Limited Reagents for directed biomarker signal amplification
US9139869B2 (en) 2010-01-19 2015-09-22 Sirigen Inc. Reagents for directed biomarker signal amplification
US10962546B2 (en) 2010-01-19 2021-03-30 Sirigen Ii Limited Reagents for directed biomarker signal amplification
US10955417B2 (en) 2010-01-19 2021-03-23 Sirigen Ii Limited Reagents for direct biomarker signal amplification
US9547008B2 (en) 2010-01-19 2017-01-17 Sirigen Inc. Reagents for directed biomarker signal amplification
US10092000B2 (en) 2010-07-13 2018-10-09 Stc.Unm Structure, synthesis, and applications for oligo phenylene ethynylenes (OPEs)
US10174042B2 (en) 2010-07-13 2019-01-08 Stc.Unm Structure, synthesis, and applications for poly (phenylene) ethynylenes (PPEs)
US9527806B2 (en) 2010-07-13 2016-12-27 Stc.Unm Structure, synthesis, and applications for poly (phenylene) ethynylenes (PPEs)
US10750746B2 (en) 2010-07-13 2020-08-25 University Of Florida Research Foundation, Inc. Structure, synthesis, and applications for poly (phenylene) ethynylenes (PPEs)
US10058099B2 (en) 2011-08-03 2018-08-28 Stc.Unm Antimicrobial materials and methods
US9549549B2 (en) 2011-08-03 2017-01-24 Stc.Unm Antimicrobial materials and methods
WO2015054484A1 (en) * 2013-10-09 2015-04-16 The University Of Akron Integrated zwitterionic conjugated polymers for bioelectronics, biosensing, regenerative medicine, and energy applications
US9695275B2 (en) 2013-10-09 2017-07-04 The University Of Akron Integrated zwitterionic conjugated polymers for bioelectronics, biosensing, regenerative medicine, and energy applications
US10407537B2 (en) 2013-10-09 2019-09-10 The University Of Akron Integrated zwitterionic conjugated polymers for bioelectronics, biosensing, regenerative medicine, and energy applications
US9968698B2 (en) 2013-11-08 2018-05-15 Stc. Unm Charged singlet-oxygen sensitizers and oppositely-charged surfactants
US10533991B2 (en) 2014-03-14 2020-01-14 Stc.Unm P-phenylene ethynylene compounds as bioactive and detection agents
US9750250B2 (en) 2015-01-14 2017-09-05 Stc.Unm Conjugated polyelectrolytes and methods of using the same
US10638759B2 (en) 2015-01-14 2020-05-05 University Of Florida Research Foundation, Inc. Conjugated polyelectrolytes and methods of using the same
US11179502B2 (en) * 2015-06-12 2021-11-23 University of Pittsburgh—of the Commonwealth System of Higher Education Biomimetic hydrogel scaffolds and related methods
US10307513B2 (en) * 2015-06-12 2019-06-04 University of Pittsburgh—of the Commonwealth System of Higher Education Biomimetic hydrogel scaffolds and related methods
US10772851B2 (en) 2017-02-03 2020-09-15 Aaron Kurt Neumann Treatment and prevention of fungal infections
WO2024161291A1 (en) * 2023-01-30 2024-08-08 Asima Health Inc. Impedance-based liquid biopsy system and method for detecting and screening cancer

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