| Polyelectrolyte complex(e.g.zwitterionic polythiophenes) with a receptor (e.g.polynucleotide, antibody etc.) for biosensor applications -> Monitor Keywords |
|
|
  Polyelectrolyte complex(e.g.zwitterionic polythiophenes) with a receptor (e.g.polynucleotide, antibody etc.) for biosensor applicationsUSPTO Application #: 20060175193Title: Polyelectrolyte complex(e.g.zwitterionic polythiophenes) with a receptor (e.g.polynucleotide, antibody etc.) for biosensor applications Abstract: A complex between a conjugated polyelectrolyte, and one or more receptor molecules specific for a target biomolecule analyte, the polyelectrolyte and the receptor being non-covalently bound to each other, is usable as a probe for biomolecular interactions. It also relates to a method of determining selected properties of biomolecules. Thereby, a complex as above is exposed to a target biomolecule analyte whereby the analyte and the receptor interact, and a change of a property of the polyelectrolyte in response to the interaction between the receptor and the analyte is detected. The detected change is used to determine the selected property of the biomolecule. (end of abstract) Agent: Young & Thompson - Arlington, VA, US Inventors: Olle Inganas, Peter Asborg, Peter Nilsson USPTO Applicaton #: 20060175193 - Class: 204242000 (USPTO) Related Patent Categories: Chemistry: Electrical And Wave Energy, Apparatus, Electrolytic, Cells The Patent Description & Claims data below is from USPTO Patent Application 20060175193. Brief Patent Description - Full Patent Description - Patent Application Claims
FIELD OF THE INVENTION [0001] 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. BACKGROUND [0002] The development of materials that are capable of selectively detecting biomolecular interactions have come under increasing attention, owing to their large potential for molecular electronics and biosensors. In this regard, conjugated polymers (CPs) 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. This has been accomplished by making conjugated (and sometimes luminescent) 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. [0003] The physical and chemical properties of 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. However, these methods, which use polyanionic or polycationic conjugated polyelectrolytes, based on interactions mainly dominated by electrostatic forces, sometimes requires labelling of the analyte. Methods without any labelling of the analyte or any covalent attachment of the receptor would be attractive. SUMMARY OF THE INVENTION [0004] Thus, there remains a need for simpler and more sensitive methods for detection of molecular interactions. Methods based on conjugated polyelectrolytes that can create versatile interactions with molecules and detect molecular interactions, without any labelling of the analytes or any covalent attachment of the receptors, would therefore be desirable. [0005] The object of the present invention is therefore to provide means and methods that meet these and other needs. [0006] 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. [0007] For the purpose of this invention, 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. [0008] Suitably 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. [0009] In a further aspect of the invention, there is provided 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. In still another aspect of the invention there is provided 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. [0010] 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. BRIEF DESCRIPTION OF THE DRAWINGS [0011] 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. [0012] FIG. 2 schematically illustrates the method according to the invention. [0013] FIG. 3 shows the absorptionspectra of 1.16 .mu.mol POWT (on a monomer basis) and 0 mol (.quadrature.), 6.4 nmol (.diamond.) 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 (.DELTA.). [0014] FIG. 4 shows the emission spectra of 23.1 nmol POWT (on a monomer basis) and 0 mol (.quadrature.), 1.28 nmol (.diamond.), 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 (.DELTA.). All of the emission spectra were recorded with excitation at 400 nm. [0015] 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 (.diamond.), 1.0 equivalent of a negatively charged peptide, JR2E (.DELTA.), 0.5 equivalents JR2E plus an addition of 0.5 equivalents JR2K (.quadrature.), 0.5 equivalents JR2K plus an addition of 0.5 equivalents JR2E (.box-solid.), 2.0 equivalent JR2K (.diamond-solid.), and 2.0 equivalent JR2E (.tangle-solidup.) 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. [0016] FIG. 6 shows the Emission spectra of 26.1 nmol POWT in 20 mM Na-phosphate pH 7.5, upon addition of 4.9 .mu.M of a synthetic peptide, with a receptor site for carbonic anhydrase (thin line), and after addition of 13 .mu.M of carbonic anhydrase (bold line). [0017] 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.times.100 .mu.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). [0018] 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. [0019] FIG. 9 shows the microcontact printing of POWT. A square net of POWT on plasma etched polystyrene, with lines 25 .mu.m wide surrounding the polystyrene squares of 100.times.100 .mu.m. Optical microscopy in reflected light. [0020] 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. Continue reading... Full patent description for Polyelectrolyte complex(e.g.zwitterionic polythiophenes) with a receptor (e.g.polynucleotide, antibody etc.) for biosensor applications Brief Patent Description - Full Patent Description - Patent Application Claims Click on the above for other options relating to this Polyelectrolyte complex(e.g.zwitterionic polythiophenes) with a receptor (e.g.polynucleotide, antibody etc.) for biosensor applications patent application. ###  How KEYWORD MONITOR works... a FREE service from FreshPatents1. Sign up (takes 30 seconds). 2. Fill in the keywords to be monitored. 3. Each week you receive an email with patent applications related to your keywords. Start now! - Receive info on patent apps like Polyelectrolyte complex(e.g.zwitterionic polythiophenes) with a receptor (e.g.polynucleotide, antibody etc.) for biosensor applications or other areas of interest. ### Previous Patent Application: Optoelectronic probe Next Patent Application: Electrochemical oxygen generator module assembly Industry Class: Chemistry: electrical and wave energy ### FreshPatents.com Support Thank you for viewing the Polyelectrolyte complex(e.g.zwitterionic polythiophenes) with a receptor (e.g.polynucleotide, antibody etc.) for biosensor applications patent info. IP-related news and info Results in 8.8203 seconds Other interesting Feshpatents.com categories: Tyco , Unilever , Warner-lambert , 3m |
||
