MXPA00002062A - Sol-gel matrices for direct colorimetric detection of analytes - Google Patents

Abstract

The present invention relates to methods and compositions for the direct detection of analytes using color changes that occur in immobilized biopolymeric material in response to selective binding of analytes to their surface. In particular, the present invention provides methods and compositions related to the encapsulation of biopolymeric material into metal oxide glass using the sol-gel method.

Description

SUN GEL MATRICES FOR DIRECT COLORIMETRIC DETECTION OF ANALYZES D E S C R I P C I N FIELD OF THE INVENTION The present invention relates to methods and compositions for the direct detection of analytes using color changes that occur in the immobilized biopolymer material in response to selective binding of analytes to its surface. ? BACKGROUND OF THE INVENTION A main goal of the investigation in the detection of analytes is to develop economic, rapid, reliable and sensitive detectors. Unfortunately, the technologies developed to date have met only some of these goals and no single device has sufficiently achieved a majority of these goals. classical detection methods such as liquid chromatography (LC), gas chromatography (GC) and supercritical fluid chromatography (SFC) combined with mass spectrometry, have been widely used and provide accurate identification of analytes and quantitative data. However, these techniques take time, are extremely expensive, require preconcentration of the sample and are also difficult or impossible to adapt to use in the field. Bioperceptors (ie, devices containing biological material attached to a transducer apparatus) have been developed to overcome some of the disadvantages of classical analyte detection techniques. Many of the biopreceptors normally used are associated with transducer devices that use photometry, fluorimetry and chemiluminescence; optical fibers and optical direct detection (eg grating coupler; surface plasmon resonance; potentiometric and amperometric electrodes, field effect transistors, piezoelectric preceptors, and surface acoustic wave (Kramer, J. AOAC Intern 79: 1245 [. 1996]). However, there are drawbacks over these devices, including their dependence on a transducer device, which prevents miniaturization and requires a power source. these disadvantages make these very complex, costly or unmanageable devices for many applications detection of routine analytes, such as field work or home use Additionally, many of these devices are limited by the lack of stability and availability of biological materials (eg, proteins, antibodies, cells and organelles). Immunity testing methods are also used to detect certain types of analytes. The antibodies are developed to bind specifically to an objective of interest (for example an analyte). By distinguishing the antibody (for example with a dye or a fluorescent or radioactive material), the ligand or binding of an antibody to an analyte can be detected. However, immunity testing methods are limited because they require the production of antibodies against each analyte of interest. Antibodies can not be generated against some types of analytes and their generation can take time and be expensive. The technique remains with the need for analyte detectors that provide the specification of bioperceptors but that also provide cost-efficiency, stability, accuracy, reliability, reproducibility and robustness, ie lack of available technologies. In particular, the development of devices that can be miniaturized with controlled forms and that are not based on a source of energy, would also be very beneficial, particularly for routine field work and home use.

THE INVENTION The present invention relates to methods and compositions for the direct detection of analytes using color changes that occur in immobilized biopolymer material in response to selective binding of analytes to its surface. The present invention provides various methods and compositions useful for the detection of analytes. In one embodiment, the present invention provides methods for immobilizing biopolymer material, comprising: providing a metal oxide, a biopolymeric material, an acid, a buffer, and sonification means; sonicate the metal oxide and the acid using the sonification means to produce a sonified solution; add the buffer to the sonicated solution to produce a buffered solution; and adding the biopolymer material to the buffered solution to produce an organic / inorganic solution. In alternative embodiments of the methods, the present invention further comprises the steps of applying the organic / inorganic solution to a forming support to produce a formed organic / inorganic solution; and gelling the organic / inorganic solution formed to produce an organic / inorganic device. In preferred embodiments, the metal oxide comprises tetramethylorthosilicate, although it is contemplated that any material that can be employed to produce a substantially transparent porous glass may be used in the methods of the present invention. In some embodiments, the biopolymeric material is selected from the group consisting of liposomes, films, multilayers, braided shapes, helical lamellae, tubular and fiber-shaped configurations, solvated rods, solvated coils and combinations thereof. In other embodiments, the biopolymer material comprises a plurality of self-assembling monomers selected from the group consisting of diacetylenes, acetylenes, alkenes, thiophenes, polythiophenes, imides, acrylamides, methacrylates, vinyl ether, malic anhydride, urethanes, allylamines, siloxanes, anilines, pyrroles , vinylpyridinium and combinations thereof, although any self-assembling monomer that can form biopolymeric material, is contemplated by the present invention. In the preferred embodiments, the diacetylenes are selected from the group consisting of 5,7-docosadiyanoic acid, 10, 12-pentacosadiynoic acid, 5,7-pentacosadiynoic acid and combinations thereof. In still other embodiments, the self-assembling monomers contain front groups selected from the group consisting of carboxylic acid, hydroxyl groups, amine groups, amino acid derivatives and hydrophobic groups, although any frontal group that exists or can be synthesized in self-assembling monomers, has been contemplated by the invention claimed herein. In some preferred embodiments, the biopolymer material further comprises a ligand. In some embodiments, the ligand is selected from the group consisting of peptides, carbohydrates, nucleic acids, biotin, drugs, chromophores, antigens, chelation compounds, molecular recognition complexes, ionic groups, polymerizable groups, linker groups, donor electrons, electron-accepting groups, hydrophobic groups, hydrophilic groups, receptor-binding groups, antibodies and combinations thereof, although any ligand that can be linked to or associated with biopolymeric material is contemplated by the present invention. In some preferred embodiments, the acid comprises hydrochloric acid, while in other preferred embodiments, the buffer comprises 3- [N-morpholino] propanesulfonic acid. In other embodiments, the sonification is conducted at a temperature of 0 ° C to 20 ° C. The present invention further provides an organic / inorganic device produced in accordance with any and all of the methods described above. In addition, the present invention provides biopolymer material encapsulated in sol-gel glass. In some embodiments, the glass comprises tetramethyl orthosilicate. In some preferred embodiments, the sol-gel glass encapsulated biopolymer material is selected from the group consisting of liposomes, films, multilayers, trefoil, laminar, helical, tubular, and fiber-shaped configurations, solvated rods, solvated coils and combinations thereof. In particularly preferred embodiments, the biopolymer material comprises self-assembling monomers selected from the group consisting of diacetylenes, acetylenes, alkenes, thiophenes, polythiophenes, imides, acrylamides, methacrylates, vinyl ether, malic anhydride, urethanes, allylamines, siloxanes, anilines, pyrroles, vinylpyridinium and combinations thereof, although any self-assembling monomer that can form biopolymeric material is contemplated by the present invention. In preferred embodiments, the diacetylenes are selected from the group consisting of 5,7-docosadiyanoic acid, 10, 12-pentacosadiynoic acid, 5,7-pentacosadiynoic acid and combinations thereof. In still other embodiments, the self-assembling monomers contain front groups selected from the group consisting of carboxylic acid, hydroxyl groups, amine groups, amino acid derivatives and hydrophobic groups, although any frontal group that exists or can be synthesized in self-assembling monomers, has been contemplated by the invention claimed herein. In some preferred embodiments, the sol-gel glass-encapsulated biopolymer material further comprises a ligand. In some embodiments, the ligand is selected from the group consisting of peptides, carbohydrates, nucleic acids, biotin, drugs, chromophores, antigens, chelation compounds, molecular recognition complexes, ionic groups, polymerizable groups, linker groups, electron donors, electron-accepting groups, hydrophobic groups, hydrophilic groups, receptor-binding groups, antibodies and combinations thereof, although any ligand that can bind to, or associate with biopolymeric material, is contemplated by the present invention. The present invention further provides methods for detecting analytes, comprising: providing a biopolymer material encapsulated in sol-gel glass, a detection means and one or more analytes; exposing the biopolymer material encapsulated in sol-gel glass to the analyte to produce a response; and detecting the response using the detection means. In preferred embodiments, the glass comprises tetramethyl orthosilicate, although any other material that can be used to produce a porous, substantially transparent glass is contemplated to be used in the methods of the present invention. In some embodiments, the biopolymeric material is selected from the group that it consists of liposomes, films, multilayers, trensade, laminar, helical, tubular, and fiber-shaped configurations, solvated rods, solvated coils, and combinations thereof. In other embodiments, the biopolymer material comprises a plurality of self-assembling monomers selected from the group consisting of diacetylenes, acetylenes, alkenes, thiophenes, polythiophenes, imides, acrylamides, methacrylates, vinyl ether, malic anhydride, urethanes, allylamines, siloxanes, anilines, pyrroles , vinylpyridinium and combinations thereof, although any self-assembling monomer that can form biopolymeric material, is contemplated by the present invention. In the preferred embodiments, the diacetylenes are selected from the group consisting of 5,7-docosadiyanoic acid, 10, 12-pentacosadiynoic acid, 5,7-pentacosadiynoic acid and combinations thereof. In yet other embodiments, the self-assembling monomers contain front groups selected from the group consisting of carboxylic acid, hydroxyl groups, amine groups, amino acid derivatives and hydrophobic groups, although any front group that exists or can be synthesized in self-assembling monomers, has been contemplated by the invention claimed herein. In some preferred embodiments, the biopoimeric material further comprises a ligand. In some embodiments, the ligand is selected from the group consisting of peptides, carbohydrates, nucleic acids, biotin, drugs, chromophores, antigens, chelation compounds, molecular recognition complexes, ionic groups, polymerizable groups, linker groups, electron donors, electron-accepting groups, hydrophobic groups, hydrophilic groups, receptor-binding groups, antibodies and combinations thereof, although any ligand that can bind to, or associate with biopolymeric material, is contemplated by the present invention. In some embodiments, the analyte is selected from the group consisting of small molecules, pathogens, bacteria, membrane receptors, membrane fragments, enzymes, drugs, antibodies and combinations thereof, but any analyte that can be detected through the interaction with a ligand or biopolymeric material, has been contemplated by the present invention. In yet other embodiments, the sol-gel glass-encapsulated biopolymer material comprises an identity plate. In preferred embodiments, the detection means is selected from the group consisting of visual inspection, spectrometry, optical fiber, quartz oscillators, electrodes and scintillation, although any other detection means that provides analysis of the presence of an analyte, is contemplated by the present invention. In some embodiments, the response is used as a competitive binding measurement to quantify and characterize the presence of natural binding sites. In other embodiments, the biopolymer material encapsulated in sol-gel glass comprises an array.

DESCRIPTION OF THE FIGURES Figure 1 shows a diagram of a receptor-linker-ligand complex wherein compound 1 shows a group of sialic acid linked to 10, 12-pentacosadiinoic acid (compound 2) through a linking group. Figure 2 shows a visible absorption spectrum of "blue phase" DCDA liposomes, entrapped in a sol-gel glass. Figure 3 shows a visible absorption spectrum of "red phase" DCDA liposomes, entrapped in a sol-gel glass. Figure 4 shows a diagram of ADP with alterations in the position of the diacetylene group of acid 10,12 to 5,7-pentacosadiinoic acid. Figure 5 shows a visible absorption spectrum of the DCDA liposomes bound by the "blue phase" sialic acid, entrapped in a sol-gel glass. Figure 6 shows a visible absorption spectrum of the DCDA liposomes bound by the "red phase" sialic acid, entrapped in a sol-gel glass.

Figure 7 shows a representation of the porous structure of the material prepared with sol-gel, acting as a selective size barrier. Figure 8 shows a visible absorption spectrum of diacetylene material exposed to 1-octanol. Figure 9 shows a bar graph indicating the colorimetric responses of PDA material to several VOCs and a table showing the concentration of VOCs. Figure 10 shows a graph comparing the colorimetric responses of 1-butanol with the concentration of 1-butanol. Figure 11 shows a visible absorption spectrum of PDA bound with acid before (solid line) and after (interrupted lines) of exposure to influenza virus for: A) blue / pink form; and (B) purple / orange material. Figure 12 shows a visible absorption spectrum of PDA bound with sialic acid, (A) before; and (B) after exposure to cholera toxin. Figure 13 shows PDA leads for use in detection arrays. Figure 14 shows the organic synthesis of compound 2.10.

GENERAL DESCRIPTION OF THE INVENTION The present invention relates to methods and compositions for the direct detection of analytes using color changes in immobilized biopolymer material that occur in response to selective binding of analytes to their surface. In particular, the present invention provides methods and compositions related to polymerized biological materials immobilized in porous glass that undergo conformational changes when exposed to analytes, producing a detectable color change, although other means of immobilization have also been contemplated. The present invention provides direct detection of the presence of a wide scale of analytes by color changes, including, but not limited to, small molecules, pathogens, bacteria, membrane receptors, membrane fragments, volatile organic compounds (VOCs), enzymes, drugs, antibodies and other relevant materials. The results can be interpreted by an untrained observer and the methods can be conducted under environmental conditions, making them manageable for numerous uses, including, but not limited to, home test diagnostics, detection of pathogens that are created in the air or in the air. water for military applications, doctors office or careful testing and many other applications. The present invention provides analyte detection technology that does not require a power source and is cost efficient, stable, accurate, reliable, consistent and robust and that can be produced in a variety of configurations and dimensions. These improved qualities provide an ideal basis for use in conjunction with optical fiber methods for remote sensing, examination of compound deposits (eg drug testing), drug testing, water supply testing and any area where a Exact colorimetric examination. Recent research has found that liposomes and other lipid-based materials can serve as optical sensory sensors for virus detection (see, for example, Reichert et al., J. Am. Chem. Soc. 117: 829 [1995]; Spevak et al. , J. Am. Chem. Soc. 115: 1146

[1993]; and Charych et al. , (Science 261: 585 [1993]). These materials present fast response times, selectively and optical signals that are easily monitored. As the aggregates float freely in solution, these lipid-based detectors show predisposition as simple test systems. The present invention provides embodiments in which these materials are used immobilized in sol-gel glass, which offer the advantages of additional chemical and physical stabilization of the material, allow for easy handling and the opportunity for recovery and reuse. To date, this effort has been frustrated by the difficulty to immobilize lipid assemblies to surface. A few methods have been developed that overcome some of the difficulties, using polysaccharides and biocompatible acrylate gels to encapsulate liposomes. However, until the development of the present invention, under volume of entrapment of liposomes, the inability to immobilize immobilized preformed liposomes and material instability at elevated temperatures, have still been inconvenient to solve. The present invention provides a means for immobilizing preformed liposomes and other biopolymer material at low temperature (e.g., 4 ° C) to room temperature, with high entrapment volume of material, in a porous, solid metal oxide gel matrix. using the sol-gel method (See generally, Brinker and Scherer, Sol-Gel Science, Academic Press, San Diego [1995]). Before the present invention, the occlusion or entrapment of lipids in sol-gel material has not been previously reported. The unique properties of sol-gel materials of the present invention such as optical transparency, durability and aggregate properties (eg porosity, surface functionality, thin films and volumetric materials) provide an ideal material for sensor applications. Sol-gel encapsulation not only provides an excellent way to immobilize liposomes and other biopolymer materials, but also the optical clarity of the metal oxide gel makes it ideal for optical sensing applications. This unique compound can be easily applied to surfaces and molded into any desired shape, allowing most of the platforms of any sensor to be configured. The robust nature of the sol-gel material converts the biopolymer-based materials into sensing materials that achieve good portability, handling, durability and life to improved storage (this is shelf life) while maintaining its sensitivity. In addition, the porous structure of the metal oxide gel and the ionic surface can be added to provide a primary analysis mechanism and a preconcentrator for selective recognition and detection of target analytes. The biopolymer / sol-gel material is a unique class of an organic-inorganic compound that offers high matrix stability against microbial attack, temperature changes and physical stresses as opposed to polysaccharide and acrylate gels. The ambient temperature at which the formation of the gel and the biologically inert metal oxide matrix is made allows a wide scale of biopolymer material and biopolymer material trapped in protein to be immobilized. Accordingly, the present invention provides methods and compositions that meet many of the goals of the analyte detection field and overcome many of the disadvantages of commonly available technologies (e.g., classical methods, bioperceptors, and immunity tests. invention provides significant advantages over the previously used bioperceivers, since the embodiments of the present invention do not depend on the transducer technologies.Many of the proposed bioperceptors, can not be used because of the difficulties in transducing the molecule recognition event into a measurable signal. In addition, the transducers of the normally developed devices add costs, create the requirement of a power source, are more difficult to use by untrained personnel and are limited in terms of miniaturization and portability. Also, many bioperceptors do not exhibit the long term stability and robustness of the invention claimed herein. The immunity tests are much more limited in the scale of analytes that can detect and do not characterize the stability and robustness of the invention claimed herein. The constructions and inventive methods can test very small biological molecules or other molecules for which antibodies can not be developed. These objective materials may include organic solvents or contaminants present at extremely low levels.

DEFINITIONS To facilitate understanding of the present invention, a number of terms and phrases are defined below: As used herein, the term "immobilization" refers to the attack or entrapment, whether chemically or otherwise, of a material to another material (for example, a solid support), in a way that restricts the movement of the material.

As used herein, the term "material" and "materials" refer in their broadest sense to any composition of matter. As used herein, the term "biopolymeric material" refers to composites of polymerized biological molecules (e.g., lipids, proteins, carbohydrates, and combinations thereof). These materials include, but are not limited to, films, ampoules, liposomes, multilayers, aggregates, membranes, and solvated polymers (eg, polythiophene aggregates such as sticks, and solvent coils). As used herein, the term "biopolymer films" refers to polymerized organic films, which are used in a thin section or in the form of a layer. Those films may include, but are not limited to monolayers and bilayers. Biopolymer films can mimic biological cell membranes, (for example in its ability to interact with other molecules such as proteins or analytes). As used herein, the term "sol-gel" refers to composite preparations of porous metal oxide glass structures. These structures may have biological material or other material trapped inside the porous structures. The phrase "sol-gel matrices" refers to the structures comprising the porous metal oxide glass, with or without entrapped material. The term "sol-gel material" refers to any material prepared by the sol-gel process that includes the glass material itself and any material trapped within the porous structure of the glass. As used herein, the term "sol-gel method" refers to any method that results in the production of porous metal oxide glass. In some embodiments the term "sol-gel method" refers to those methods conducted under conditions of moderate temperatures. The term "sol-gel glass" and "metal oxide glass" refers to glass material prepared by the sol-gel method and includes inorganic material or mixed organic / inorganic material. These materials used to produce the glass may include, but are not limited to, aluminates, aluminosilicates, titanates, ormosils (organically modified silanes) and other metal oxides. As used herein, the term "direct colorimetric detection" refers to the detection of color changes, without the aid of an intervention processing step (e.g., conversion of a color change into an electronic signal, which is processed by an interpretation device). It is intended for the term to encompass visual observation (for example, observation with the human eye). As used herein, the term "analytes" refers to any material that is to be analyzed. Such material may include, but is not limited to, molecules, bacteria, compounds, viruses, cells, antibodies and parts of cells. As used herein, the term "selective binding" refers to the attachment of a material to another material in a dependent manner under the presence of a particular molecular structure (ie, specific binding). For example, a receptor will selectively bind ligands containing the chemical structures complementary to the site or the binding sites of the ligands. As used herein, the term "bioperceptors" refers to any sensing device that is partially or wholly composed of biological molecules. In a traditional sense, the term refers to an "analytical system or tool consisting of an immobilized biological material (such as an enzyme, antibody, whole cell, organelle, or combinations thereof) in intimate contact with a transducer device that will convert the biochemical signal in a quantifiable electrical signal "(Gronow, Trends Biochem, Sci. 9: 336 [1984]).

As used herein, the term "transducer device" refers to a device that is capable of converting a non-electrical phenomenon into electrical information and transmitting the information to a device that interprets the electrical signal. These devices may include, but are not limited to, devices that use photometry, fluorimetry and chemiluminescence; fiber optics and direct optical detection (for example, grid coupler); plasmon surface resonance; potentiometer and ammeter electrodes; field effect transistors; piezoelectric sensors; and surface acoustic waves. As used herein, the term "miniaturization" refers to a reduction in size, such as the size of a sample to increase its usefulness (e.g., portability, ease of handling, and ease of incorporation into arrays). As used herein, the term "stability" refers to the ability of a material to withstand deterioration or displacement and to provide reliability and dependability. As used herein, the term "conformational change" refers to the alteration of the molecular structure of a substance. It is intended for the term to encompass the alteration of the structure of a single molecule or a molecular aggregate (for example, the change of structure of polydiacetylene under the interaction with an analyte). As used herein, the term "small molecules" refers to any molecule with low molecular weight (ie, less than 10,000 atomic units of mass and preferably less than 5,000 atomic units of mass) that binds ligands, interacts with Ligands or interacts with biopolymeric material in a way that creates a conformational change. As used herein, the term "pathogen" refers to organisms that cause diseases, microorganisms or agents that include, but are not limited to, viruses, bacteria, parasites (including, but not limited to, organisms with the Protozoa phyla, Platyhelminthes, Aschelminithes, Acanthocephala and Arthropoda) fungi and prions. As used herein, the term "bacteria" and "bacteria" refers to all prokaryotic organisms, including those within all phyla in the prokaryotic kingdom. It is intended for that term to encompass all microorganisms that are considered to be bacteria, including Mycoplasma, Chlamydia, Actinomyces Streptomyces and Rickettsia. Included within this definition are all forms of bacteria including cocci bacteria, bacilli, spirochetes, spheroplasts, protoplasts, etc. The organisms "Gram-negative" and "Gram-positive" refer to patterns of discoloration, obtained with the process of Gram discoloration that is well known in the art (see for example, Finegold and Martin, Diagnostic Microbiology, 6th Edition (1982), CV Mosby St. Louis, pp. 13-15). As used herein, the term "membrane" refers to, in its broadest sense, a thin sheet or layer of material. It is intended for the term to encompass all "biomembranes" (ie, any organic membrane). , including, but not limited to, plasma membranes, nuclear membranes, organelle membranes and synthetic membranes). Typically, the membranes are composed of lipids, proteins, glycolipids, steroids, sterol and / or other components. As used herein, the term "membrane fragment" refers to any portion or part of a membrane. The term "polymerized membrane" refers to membranes that have undergone partial or complete polymerization. As used herein, the term "polymerization" encompasses any process that results in the conversion of small molecular monomers to larger molecules, which consist of repeating units. As used herein, the term "membrane receptors" refers to membrane constituents that are capable of interacting with other molecules or materials. These constituents can include, but are not limited to proteins, lipids, carbohydrates and combinations thereof. As used herein, the term "volatile organic compound" or "VOC" refers to organic compounds that are reactive (i.e., rapidly evaporate, explosive, corrosive, etc.), and typically are hazardous to human health or the environment above certain concentrations. Examples of VOCs include, but are not limited to alcohols, benzenes, toluenes, chloroforms and cyclohexanes. As used herein the term "enzyme" refers to molecules or aggregates of molecules that are responsible for catalyzing chemical and biological reactions. These molecules are typically proteins, but they can also be short peptides, RNAs or other molecules. As used herein, the term "drug" refers to a substance or substances that are used for diagnosis, treatment or prevention of diseases or conditions. Drugs act alternately on the physiology of a living organism, tissue, cell or an in vi tro system that are exposed. The term is intended to encompass antimicrobial organisms, including, but not limited to, antibacterial, antifungal and antiviral compounds. It is also intended for the term to encompass antibiotics, including those that occur naturally and synthetics and compounds produced by recombinant DNA technology. As used herein, the term "peptide" refers to any substance composed of two or more amino acids. As used herein, the term "carbohydrates" (carbohydrates) refers to a class of molecules including, but not limited to, sugars, starches, cellulose, sprouts, glycogen and similar structures. Carbohydrates can occur as components of glycolipids and glycoproteins. As used herein, the term "chromophore" refers to molecules or groups of molecules that respond to the color of a compound, material or sample. As used herein, the term "antigen" refers to any molecule or group of molecules that is recognized by at least one antibody. By definition, an antigen should contain at least one epitope (ie, the specific biochemical unit capable of being recognized by the antibody). The term "immunogen" refers to any molecule, compound or aggregate that induces the production of antibodies. By definition, an immunogen should contain at least one epitope (that is, the biochemical unit capable of causing an immune response). As used herein, the term "chelation compound" refers to any compound of or containing coordinated bonds that complete a closed ring structure. As used herein, the term "molecular recognition complex" refers to any molecule, group of molecules or molecular complex that is capable of recognizing (i.e. specifically interacting with) a molecule. As used herein, the term "ambient condition" refers to the conditions of the surrounding environment (for example, of the indoor or outdoor ambient temperature at which the experiment is performed). As used herein, the term "ambient temperature" refers, technically, to temperatures of approximately between 20 and 25 degrees centigrade. However, as it has been used in general, it refers to any ambient temperature within a general area in which an experiment is performed. As used herein, the term "home test or home test" and "test point test" refers to a test that occurs outside of a laboratory environment. Such proof may occur inside or outside of, for example, a private residence, a place of business, a public place or in a private place, in a vehicle, underwater, as well as next to the bed a patient. As used herein, the term "lipid" refers to a variety of compounds that are characterized by their solubility in organic solvents. These compounds include, but are not limited to fats, waxes, steroids, sterols, glycosipid glycosides (including gangliosides), phospholipids, terpenes, liposoluble vitamins, prostaglandins, carotenes and chlorophylls. As used herein, the phrase "lipid-based materials" refers to any material that contains lipids. As used herein, the term "virus" refers to any infectious agent, incapable of multiplying without a host cell (i.e., agents that are obligate parasites.

As used here, the phrase "freely floating aggregates" refers to aggregates that have not been immobilized. As used herein, the term "encapsulation" refers to the process of embracing, boxing or otherwise associating two or more materials such that the encapsulated material is immobilized within or on the encapsulating material. For example, the sol-gel process provides a means for encapsulating material in porous sol-gel glass material. As used herein, the term "trapped volume" refers to the volume of material encapsulated within a material. For example, the volume of liposome encapsulated within the porous structure of sol-gel glass material is the trapped volume. As used here, the term "optical transparency" refers to the property of matter by which matter is capable of transmitting light in such a way that light can be observed by visual light detectors. (for example, eyes and detection equipment). As used herein, the term "biologically inert" refers to a property of the material by which the material does not react chemically with biological material.

As used herein, the term "organic solvents" refers to any organic molecules capable of dissolving another substance. Examples include, but are not limited to, chloroform, alcohols, phenols and ethers. As used herein, the term "nanostructures" refers to microscopic structures, typically measured on a nanoscale. Such structures include several three-dimensional assemblies, including, but not limited to, liposomes, films, multilayers, braided, laminar, helical, tubular shapes and configurations in the form of fibers, and combinations thereof. Those structures can be, in some embodiments, solvated polymers in aggregate forms such as rods and coils. As used, herein the term "films" refers to any material deposited or used in a thin section or in the form of a layer. As used herein, the term "blister or vesicle" refers to small enclosed structures. Often, the structures are membranes composed of lipids, proteins, glycolipids, steroids, or other components associated with membranes. Blisters can be naturally generated (for example, the blisters present in the cytoplasm of cells that carry molecules and specific cell division functions) or can be synthetic (for example, liposomes). As used herein, the term "liposome" refers to artificially produced spherical lipid complexes that can be induced to secrete out of aqueous media. As used herein, the term "biopolymer liposomes" refers to liposomes that are composed entirely, or in part, of biopolymer material. As used herein, the term "tubular" refers to material comprising small hollow cylindrical structures. As used herein, the term "multilayer" refers to structures comprised of two or more monolayers. Individual monolayers can interact chemically with each other (eg, through covalent bonds, ionic interactions, van der Waals interactions, hydrogen bonds, hydrophobic or hydrophilic assemblies and stearic hindrance) to produce a film with novel properties (ie, properties that are different from those of monolayers alone). As used herein, the term "self-assembling monomers" refers to molecules that spontaneously associate to form molecular assemblies. In some sense, this may refer to surfactant molecules that associate to form molecular surfactant assemblies. As used herein, the term "surfactant molecular assemblies" refers to an assembly of surface active agents that contain chemical groups with opposite polarity, which form monolayers oriented in phase interfaces, which form micelles (colloidal particles in aggregation colloids) and which have detergent, foaming, wetting, emulsifying and dispersing properties. As used herein, the term "homopolymers" refers to materials that comprise a single type of polymerized molecular species. The phrase "mixed polymers" refers to materials that comprise two or more types of molecular polymerization species. As used herein, the term "ligands" refers to any ion, molecule, molecular group or other substance that binds any entity to form a larger complex. Examples of ligands include, but are not limited to, peptides, carbohydrates, nucleic acids, antibodies, or any organic molecules that bind to receptors. As used herein the terms "organic matrix" and "biological matrix" refer to collections of organic molecules that are assembled into a larger multi-molecular structure. These structures may include, but are not limited to, films, monolayers and bilayers. As used herein, the term "organic monolayer" refers to a thin film comprising a single layer of carbon-based molecules. In one embodiment, those monolayers may comprise polar molecules whereby all hydrophobic ends are aligned on one side of the monolayer. The term "monolayer assemblies" refers to structures comprising monolayers. . The term "organic polymer matrix" refers to organic matrices with which some or all of the molecular constituents of the matrix are polymerized. As used herein, the phrase "functionally front or front groups" refers to molecular groups present at the ends of the molecules (e.g., the carboxylic acid group at the fatty acid end). As used herein, the term "hydrophilic front or front group" refers to the ends of molecules that are substantially attracted to water through chemical interactions including, without limitation, hydrogen bonds, van der Waals forces, ionic interactions or linkages covalent As used herein, the term "hydrophobic front or front group" refers to the ends of molecules that self-associate with other hydrophobic entities, resulting in their exclusion from water. As used herein, the term "front or carboxylic acid front groups" refers to organic compounds that contain one or more carboxyl groups (-COOH) located at or near the end of a molecule. The term "carboxylic acid" includes carboxyl groups that are either free or exist as salts or esters. As used herein, the term "front or front detection groups" refers to the molecular group contained at the end of a molecule that is involved in the detection of a moiety or radical (eg, an analyte). As used here the terms "linker" or "Spacer molecule" refers to materials that link any entity to another. In one sense, a molecule or group of molecules can be the linker that is covalently bound to two or more other molecules (eg, by binding a ligand group to a self-assembling monomer) As used herein, the phrase " "polymeric assembly surface" refers to polymeric material that provides a surface for the assembly of additional material (eg, a biopolymer surface of a film or liposome that provides a surface for ligand binding and assembly.) As used herein, the phrase "color detection element" refers to material that is capable of providing colorimetric analysis (e.g., polymerized diacetylene.) As used herein, the term "formation support" refers to any device or structure that provides a physical support for the production of material In some embodiments, the training support provides a structure for layering and / or packaging imitating films As used herein, the term "10,12-pentacosadiynoic acid" refers to the compound with the following chemical formula: CH 3 - (CH 2) n -C 1 -C-C _ = C- (CH 2) 8 COOH. The term "5,7-pentacosadinoinoic acid" refers to the compound having the formula: CH 3 - (CH 2)? E-C = C-C = .C - "(CH 2) 3 -COOH As used herein, the term "diacetylene monomers" refers to simple copies of hydrocarbons containing two alkylene linkages (ie triple bonds of carbon to carbon) As used herein, the terms "common feeding trough" and "tundish of Langmuir-Blodgett" "common" refers to a device, usually made of Teflon, which is used to produce Langmuir films.The device contains a reservoir that holds an aqueous solution and mobile barriers to compress film material that are layered on the aqueous solution ( see for example, Roberts, Langmuir-Blodgett Films, Plenum, New York, [1990].) As used herein, the term "crystal morphology" refers to the configuration and structure of crystals that may include, but are not limited to, , to crystalline forms, orientation, text ura and size As used herein, the term "domain limit" refers to the boundaries of an area in which polymerized film molecules are homogeneously oriented. For example, a domain limit may be a physical structure of periodic material, regularly disposed of polydiacetylene (e.g., striations, ribs and grooves). As used herein, the term "domain size or dimension" refers to the typical length between domain boundaries. As used herein, the term "conjugated structure" refers to the structure of a polymer of p-PCA films that, on a macroscopic scale, appear in the form of physical ribs or striae. The term "polymer structure axis" refers to an imaginary line running parallel to the conjugated structure. The terms "intra-structure" and "inter-structure" refer to regions within a given polymer structure and between polymer structures, respectively. The structures create a series of lines or "linear striations" that extend over distances along the template surface. As used herein the term "binding" refers to the bond between atoms in molecules and between ions and molecules in crystals. The term "simple or single bond" refers to a bond with two electrons occupying the binding orbital. Simple links between atoms in molecular notations are represented by a simple line between two atoms (for example, Cs - C9). The term "double bond" refers to a union that shares two pairs of electrons. Double bonds are stronger than single bonds and are more reactive. The term "triple bond" refers to the sharing of three pairs of electrons. How it is used here, the term ene-ino refers to the alternative double and triple links. As used herein, the term "absorption" refers, in a sense, to the absorption of light. The light is absorbed if it is not reflected or transmitted through a sample. Samples that appear colored have selectively absorbed all wavelengths of white light, except for those that correspond to the visible colors that are seen. As used herein, the term "spectrum" refers to the distribution of light energies arranged in order of wavelength. As used herein, the term "visible spectrum" refers to the radiation of light containing wavelengths from about 360 nm to about 800 nm. As used herein, the term "ultraviolet irradiation" refers to exposure to radiation with wavelengths less than visible light (ie, less than 360 nm) but greater than that of X-rays (ie, , greater than 0.1 nm). Ultraviolet radiation has more energy than visible light and is therefore more effective in inducing photochemical reactions. As used herein, the term "chromatic transition" refers to changes in molecules or material that result in an alteration in the absorption of visible light. In some embodiments, the chromatic transition refers to changes in the light absorption of a sample, whereby there is a detectable color change associated with the transition. This detection can be carried out through various means including, without limitation, visual observation and spectrophotometry. As used herein, the term "thermochromic transition" refers to a chromatic transition that is initiated by a change in temperature. As used herein, the term "solid support" refers to a solid object or solid surface on which it is layered or holds a sample. The phrase "hydrophobic solid support" refers to a solid support that has been chemically treated or generated in a manner that attracts hydrophobic entities and repels water. As used herein, the phrase "solid sensor platforms" refers to any solid support used to immobilize sensing material. As used herein, the term "ambient film interface" refers to a film surface exposed to the environment or atmosphere (that is, not the surface that is in contact with the solid support). As used herein, the term "forming solvent" refers to any medium, although it typically refers to a volatile organic solvent, used to solubilize and distribute material to a desired location (e.g., to a surface to produce a film). or to a drying receptacle for depositing liposome material for drying). As used herein, the term "micelle" refers to a colloidal particle having a hydrophilic exterior and a hydrophobic interior. As used herein, the term "topochemical reaction" refers to reactions that occur at a specific location (eg, within a specific portion of a molecule or a reaction that only occurs when a certain molecular configuration is present). As used herein, the term "molding structure" refers to a solid support used as a template for design material in desired shapes and dimensions. As used here, the terms "arrangement" and "arrangement in pattern", refer to an arrangement of elements (ie, entities) in a material or device.

For example, combining sections of sol-gel material, which have different encapsulated biopolymer samples, in an analyte detection device, will constitute an array. As used herein, the term "interferers" refers to entities present in a sample of analytes that are not the analytes to be detected and that, preferably, will not identify a detection device, or difference of the or the analytes of interest. As used herein, the term "identity plate" refers to any portable device and that can be transported or carried for individual work in an analyte detection environment. As used herein, the term "halogenation" refers to the process of incorporation or the degree of incorporation of halogens (i.e., the elements fluorine, chlorine, bromine, iodine and astatine) into a molecule. As used herein, the term "aromaticity" refers to the presence of aromatic groups (ie, rings of six carbon atoms and derivatives thereof) in a molecule. As used herein, the phrase "water immiscible solvents" refers to solvents that do not dissolve in water in all proportions. The phrase "miscible solvents in water" refers to solvents that dissolve in water in all proportions. As they have been used here, the terms load "positive", "negative" and Switterionic "refer to molecules or groups of molecules that contain a positive, negative or zwitterionic net charge, respectively Zwitterionic entities contain both positively charged atoms, negatively charged atoms or groups whose charges are canceled As used here, the term "biological organisms" refers to any form of life based on carbon.As used herein, the term "in situ" refers to processes, events, objects or information that they are present or performed within the context of their natural environment, for example, in situ microscopy, refers to the analysis of the material in its natural form (ie, not sectioned, fixed or altered in another way.) As used here , the term "aqueous" refers to a liquid mixture containing water, among other components As used herein, the term "solid state" refers to reactions that involve one or more compounds. rigid or similar to solids. As used herein, the term "regularly packaged" refers to the periodic arrangement of molecules within a compressed film. As used herein, the term "filtration" refers to the process of separating samples from each other. In one embodiment, filtration refers to the separation of solids, liquids or gases, by the use of a membrane or a medium. As used here, the term "sample" is used in its broadest sense. In a sense it can refer to a crystallized product. • In another sense it can be referred to include a specimen or culture; On the other hand, it can be referred to include biological or environmental samples. Biological samples include blood products, such as plasma, serum and the like. Biological samples can be of animal origin, including humans, solid fluids or tissues. Samples of the environment include environmental material such as surface material, soil, water, crystals and industrial samples. These examples are not constructed as limiting the types of samples applicable to the present invention.

DETAILED DESCRIPTION OF THE INVENTION The invention claimed herein, comprises methods and compositions related to biopolymer material encapsulated in sol-gel and immobilized on other solid supports for the colorimetric detection of analytes. Biopolymeric materials include, but are not limited to, films, ampoules, tubular and multilayer structures, incorporated in sol-gel matrices. The biopolymeric materials contain polymerized self-assembly monomers that undergo conformational changes and chromatic transitions upon exposure of the analyte. The analytes interact either directly with the monomers or with ligands that are linked to or in association with the monomers. The entrapment in sol-gel provides stability, robustness and manipulation to the biopolymeric sensing material. With a certain biopolymeric material, a color transition occurs under the bond with the analyte, which can be seen by simple visual observation or, if desired, by color sensing equipment. The methods and compositions of the invention claimed herein find use in a wide range of analyte detection circumstances and are particularly responsible for situations where simple, rapid, accurate and cost-efficient detection is required. In certain embodiments, the present invention contemplates a variety of self-assembling monomers that are suitable for formation in biopolymeric materials. These monomers include, but are not limited to, acetylenes, diacetylenes (eg, 5,7-docosadiinoic acid, 5,7-pentacosadiinoic acid, and 10,12-pentacosadiinoic acid), alkenes, thiophenes, polythiophenes, imides, acrylamides, methacrylates, vinyl ether, malic anhydride, urethanes, allylamines, aniline siloxanes, pyrroles and vinylpyridinium. Lipids containing these groups can be homopolymers or mixed polymers. In addition, monomers are contemplated with a variety of frontal groups including, but not limited to, carboxylic acid, hydroxyl groups, primary amine functions, amino acid derivatives and hydrophobic groups. The ligands can be linked via a link arm to the self-assembling monomers, they can be directly linked to the monomers, they can be incorporated in the matrix during the polymerization process or they can be bound to the matrix after the polymerization (for example, by binding of ligands to the constituents of the matrix containing frontal groups that bind to the ligands). The group of ligands of the present invention may be of a wide variety of materials. The main criteria is that the ligands have an affinity for the selection analyte. Suitable ligands include, but are not limited to, peptides, carbohydrates, nucleic acids, biotin, drugs, chromophores, antigens, chelating compounds, molecular recognition comps, ionic groups, polymerizable groups, dinitrophenols, linker groups, donor groups, electrons or electron acceptors, hydrophobic groups, hydrophilic groups, antibodies or any organic molecule that binds receptors. The biopolymeric material may be composed of ligand-bound and unbound monomer combinations to optimize the desired colorimetric response (e.g., 5% bound ligand DCDA and 95% DCDA). Additionally, multiple ligands can be incorporated into a single biopolymer matrix. In some embodiments, the ligands are incorporated to detect a variety of bacteria and pathogenic organisms including, but not limited to, sialic acid to detect HIV (Wies et al., 333: 426 [1988]), influenza (White et al. ., Cell 56: 725 [1989]), chlamydia (Infect Imm 57: 2378 [1989]), reovirus, Streptococcus suis, Salmonella, Sendai virus, mumps, newcastle, myxovirus and Neisseria meningi tidis; 9-OAC sialic acid to detect coronavirus, encephalomyelitis virus and rotavirus; non-sialic acid glycoproteins for detecting cytomegalovirus (Virology 176: 337 [1990]) and measles virus (Virology 172: 386 [1989]); CD4 (Khatzman et al., Nature 312: 763 [1985]), vasoactive intestinal peptide (Sacerdote et al., J. of Neuroscience Research 18: 102 [1987]) and T peptide (Ruff et al., FEB Letters 211 : 17 [1987]) to detect HIV; epidermal development factor for detecting vaccines (Epstein et al., Natura 318: 663 [1985]), * acetylcholine receptor for detecting rabies (Lentz et al., Science 215: 182 [1982]); Cd3 complement receptor for detecting Epstein-Barr virus (Carel et al., J. Biol. Chem. 265: 12293 [1990]); beta-adrenergic receptor for detecting reovirus (Co et al., Proc. Nati. Acad. Sci. 82: 1494 [1985]); ICAM 1 (Marlin et al., Nature 344: 70 [1990]), N-CAM and Mab myelin-associated glycoprotein (Shephey et al., Proc. Nati, Acad. Sci. 85: 7743 [1988]) to detect rhinovirus; polio virus receptor for detecting poliovirus (Mendelsohn et al., Cell 56: 855 [1989]); fibroblast growth factor for detecting herpes virus (Kaner et al., Science 248: 1410 [1990]); oligomannose to detect Escherichia coli; GMI ganglioside to detect Neisseria meningi tidis; and antibodies to detect a wide variety of pathogenic organisms (e.g., Neisseria gonorroeae, V. Vulnificus, V. Parahaemolyticus, V. Cholerae and V. Alginolyticus). In some embodiments, the self-assembling monomers are not associated with ligands, but are directly assembled, polymerized and entrapped in so-gel. These biopolymer materials find use in the detection of certain classes of analytes, including, but not limited to, volatile organic compounds (VOCs).

Production of biopolymer material In some embodiments, the present invention provides biopolymeric material composed of polymerized monomers whose front groups are linked to a ligand through a linear structural linker. For example, Figure 1 provides a schematic illustration of an embodiment of the present invention. Compound 1 shows a receptor-linker ligand (sialic acid) attached to a terminal end of a spacer molecule. The second terminal end of the spacer molecule is attached to one of several monomers (10, 12-pentacosadiinoic acid, ie compound 2) which has been polymerized in order to form a chromatic detection element. These materials are then agitated while the polymerization occurs, causing the formation of the polymeric structures, including, but not limited to, films, liposomes, ampoules, tubular, multilayer structures and other nanostructures.

In some embodiments, the biopolymeric material used in the claimed invention comprises a biopolymer film. As described in Example 1, the film was prepared by forming layers from the desired matrix-forming material (e.g., self-assembling organic monomers) on a forming support. In preferred embodiments, the forming support was a Langmuir-Blodgett tundish and the material for forming the matrix was layered on an aqueous surface created by filling the tundish with an aqueous solution. Then, the material was compressed and polymerized to form a biopolymer film. In preferred embodiments, compression was conducted in a common Langmuir-Blodgett trough using movable barriers to compress the matrix-forming material. Compression was carried out until a packed packed monolayer was formed from the matrix forming material. As described in Example 1, in some embodiments, the matrix-forming material, located within the forming support, was polymerized by ultraviolet irradiation. All polymerization methods are contemplated by the present invention, and include, but are not limited to, gamma radiation, X-ray radiation and exposure to electron beams. In preferred embodiments, the biopolymer films comprise polymerized Langmuir-Schaefer films of 5,7,7-docosadiinoic acid (DCDA) linked or associated with the desired ligand, although other PCA's are contemplated, including, but not limited to, acid 5.7 -pentacosadiinóico (p-PCA) and 10,12-p-PCA. These films produce a visible color transition when exposed to the appropriate analyte. As described in example 1, the films were generated by spreading and compressing 5,7-DCDA and the desired ligand or the ligand-derivative of 5,7-DCDA (i.e., 5,7-DCDA chemically bound to a ligand ) on the surface of a forming support. In preferred embodiments, the forming support is a common Langmuir-Bloddgett tundish, with the materials layered on an aqueous surface, created by filling the tundish with an aqueous solution, although any Formation support that facilitates the spreading and compression of the film, is contemplated by the present invention. The diacetylene monomers (DA) were polymerized to polydiacetylene (p-PDA or PDA) using ultraviolet radiation. In preferred embodiments, the source of ultraviolet radiation is kept sufficiently far from the film, to avoid causing heat damage to the film. The crystalline morphology of the polymerized film can easily be observed between crosslinked polarizers in an optical microscope, although this step is not required by the present invention. For p-PCA, the domain dimension varied up to 1 mm, although it is contemplated that domains as large as about 3 mm can be developed. (see for example, Day and Lando, Macromolecules 13: 1478 [1980]). The structure or conjugated skeleton of alternately double and triple bonds, (i.e., eno-ino) that was generated, appeared at intense absorptions in the visible spectrum and led to a distinct blue appearance of the polymerized film. In certain embodiments, the visible blue films were then transferred to hydrophobized solid supports, such that the carboxylic acid front groups were exposed to the ambient film interface (Charych et al., Science 261: 585 [1993]) to undergo to further analysis, although the method of the present invention does not require this step. Typical linear striations of p-PDA films can be observed in the optical polymerization microscope. The material can also be characterized using an atomic force microscope.

In preferred embodiments, the biopolymeric material used in the claimed invention, comprises biopolymer liposomes. Liposomes were prepared using a test sonification method (New Liposomes: A Practical Approach, Oxford University Press, Oxford, pp 33 '104 [1990]), although any method that generates liposomes is contemplated. Self-assembling monomers, either alone or associated with a desired ligand, were dried to remove the forming solvents and resuspended in deionized water. The suspension was tested sonified and polymerized. The resulting liposome solution contained biopolymer liposomes. In some embodiments, the sonicated solution was polymerized by ultraviolet radiation, using a hand lamp. In preferred embodiments, the biopolymer liposomes comprise 5,7-DCDA alone or mixed with 5,7-DCDA linked to, or associated with the desired ligand, although other self-assembling monomers are contemplated including, but not limited to, 5,7 -PCA and 10,12-PCA. During polymerization, the appearance of the colored polymer provides a sensitive and simple molecular order test in the self-assembled nano structure. "Loose" structures such as micelles do not form the conjugated polymer, possibly due to the topochemical nature of the polymerization reaction. The conjugated eno-ino structure of polydiacetylene liposomes resulted in the appearance of an intense blue / purple solution. In other embodiments, it is contemplated that variations in the heating and cooling rates in the agitation methods and in the materials of the biopolymer material will provide other nanostructures. These nanostructures include, but are not limited to, multilayers, braided, laminar, helical, tubular structures and fiber-like configurations and combinations thereof. These structures can be, in some embodiments, solvated polymers in such aggregated forms, such as rods and coils. For example, it has been shown that the chain length of the monomers effect the type of aggregate they form in solution (Okahata and Kunitake, J. Am. Cheem. Soc. 101: 5231 [1979]).

Entrapment of biopolymeric material by the Sol-Gel method.

Whereas the sol-gel process has been used to trap organic molecules such as dyes and biomolecules in silica gelatins (See, for example, Avnir, Accounts Chem. Res. 28: 328 [1995]; Yamanaka et al., Am. Chem. Soc. 117: 9095 [1995], Miller et al., Non-Cryst, Solids 202: 279 [1996], and Dave et al., Anal. Chem. 66: 1120a [1994]), before development of. In the present invention, immobilization in self-organized molecular aggregates (eg, biopolymeric material, aggregates of self-assembled monomers and liposomes) was not carried out in sol-gel materials. Embodiments of the claimed invention herein successfully provide for the immobilization of spherical, two-layered lipid aggregates, and liposomes using an aqueous sol-gel process. These molecular structures and in particular the liposomes, biological lipid compounds or biomimetics / that is, natural mimics) are very robust under aqueous conditions and environmental temperatures but can easily be degraded in the presence of organic solvents and high temperatures. The sol-gel process provides an easy method to immobilize molecular aggregates with undetectable structure modification, creating robust structures that are easily manufactured in any desired size shapes. The silica sol-gel material was prepared by sonating tetramethyl orthosilicate, water and hydrochloric acid under cooling conditions until a single-phase solution was obtained. The use of metal oxides, other than the tetramethyl orthosilicate, is contemplated by the present invention, both as facilitating entrapment and formation of substantially transparent glass material. Those metal oxides include, but they are not limited, silicates, titanates, aluminates, ormosils and others. This compound was emptied into a desired molding structure and allowed to gelatinize at room temperature. The present invention is not intended to be limited by the type of molding structure employed, since it is contemplated that a variety of structures may be applied to generate gelatins of any desired shape and configuration including, without limitation, buckets, flat surfaces to generate Thin films, plastic, ceramic or molded metal to generate identity plates, etc. The present invention is not intended to be limited to gelatinization at ambient temperatures, since any temperature scale that facilitates the production of functional analyte detection gels has been contemplated. In one embodiment, DCDA liposomes were incorporated into the sol-gel glass, although incorporation of any biopolymer structure is contemplated by the present invention. After the sol-gel procedure as described above, gelatinization occurs in a few minutes, producing gels with a violet color. The visible absorption spectra of the polydiacetylene liposomes, as shown in Figure 2, remained unchanged in the sol-gel matrix compared to the liposomes in solution. After heating the liposomes at 55 ° C, a thermochromic transition from blue to red occurred which is characteristic of the polydiacetylene materials. The blue to red phase materials remained similarly unchanged in the sol-gel state as compared to solution as shown in the spectrum of Figure 3. Accordingly, the invention claimed herein provides a sun matrix -gel that is compatible with the most fragile of biopolymer structures (ie, liposomes) and maintains those physical properties that were observed in volumetric solution.

Colorimetric detection.

Sol-gel materials containing biopolymer material can be used as detectors for colorimetric analytes. Various spectral changes of trapped biopolymer materials can be used to detect the presence or absence of target analytes. Spectral signal amplification means, well known in the art, such as scintillators, can also be used to detect analyte levels. Because of the nature of the signal, it has been contemplated that detection methods can be automated, if desired. However, automation is not required to practice the present invention. In preferred embodiments of the claimed invention, a color shift was observed simply by visual observation. Therefore, the present invention can be easily used by any untrained observer, such as a user in a home. In alternative embodiments, spectral analysis equipment well known in the art is employed to detect changes in spectral qualities beyond the limits of simple visual observation, including optical density at a particular light wavelength of illumination. For example, using a spectrometer, the spectrum of the material was measured before and after the introduction of analytes and the colorimetric response (% CR) was measured. The visible absorption spectrum of the material before the exposure of the analytes was measured as B0 = Ix / (Iy + Ix), where "B" represents the percentage of a given color phase at a wavelength of Ix compared at a reference wavelength Iy. Then, the spectrum was taken after exposure to the analyte and a similar calculation was made to determine Bfinai. The colorimetric response was calculated as% CR = [(Bo - Bfinai) / B0] x 100%. Additionally, the invention claimed herein, if desired, may be linked to a transducer device. The association of self-assembled monomeric materials with transducers has already been described using optical fibers (see, for example, Beswick and Pitt, J. Colloid Interface Sci. 124: 146 [1988]) and Zhao and Reic ert, Langmuir 8: 2785 [ 1992]), quartz oscillators (See, for example, Furuki and Pu, Thin Solid Films 210: 471 [1992], and Kepler et al., Anal. Chem. 64: 3191 [1992]); or electrode surfaces (see, for example, Miyasaka et al., Chem. Lett., p 627 [1990] and Bilewicz and Majda, Langmuir 7: 2794 [1991]).

However, unlike these examples, the present invention provides a uniquely stable robust material that is easily associated with a transducer device. Additionally, the embodiments of the present invention provide double verification by observing the color change in the material.

Detection of analytes The biopolymer materials trapped in sol-gel created by the methods of the invention claimed herein, can be used to detect a wide variety of analytes, including, but not limited to, small molecules, pathogenic organisms, bacteria, membrane receptors, fragments of membranes, volatile organic compounds (VOCs), enzymes, drugs, antibodies and other relevant materials by observing color changes that occur in analyte linkers. The invention claimed herein, provides very mild test conditions, providing the ability to detect small biomolecules in a closely natural state and avoids the risks associated with the modification or degradation of the analyte.

Certain embodiments of the invention claimed herein, contemplates the generation of a large palette of polymerizable lipids with different chemicals from front groups within a single device to increase selectivity, an important factor in the present invention. In one embodiment, lipid-polymer perceptors trapped in sol-gel can be employed using an array format (or an "optical nose")., by analogy with mammalian nose pattern recognition capabilities). By using the array format, several advantages can be achieved that overcome the drawbacks of a single-preceptor implementation. This includes the ability to use partially selective sensors and measure multicomponent samples. The sensitivities of a given lipid to a given solvent can be determined in order to generate identifiable fingerprint characteristics of each solvent. For example, the lipid-polymer film of a derivative A of p-PDA can completely convert to an orange phase in the presence of benzene (% CR = 100), while a derivative B of p-PDA can have a% CR of 70 giving rise to a pink color, and a C derivative of p-PDA may have a% CR of 40 producing a purple color and a D derivative of p-PDA may have no change (ie, therefore it remains blue / purple). The response of an orange / pink / purple / blue-purple trace will indicate the presence of benzene. Clearly, the higher the number of elements in the array, the greater the chance of positive identification for a given analyte. The use of biopolymer material trapped in sol-gel, facilitates the generation of those arrangements, since you can create sections of gel of any size and shape desired and incorporate in a small device, easily readable and interpretable. In other embodiments, arrays of biopolymer material may be immobilized on a variety of solid supports, including, but not limited to, cellulose, nitrocellulose, and filter paper. For example, the liposomes that are incorporated in the present invention have been loaded into an ink cartridge of a jet ink printer and used to print biopolymer liposome material onto a paper as it is considered to be ink. The liposome material present on the paper maintains its colorimetric properties. This embodiment demonstrates the ease with which patterns can be generated in any desired shape and location. By using multiple cartridges (for example, using a color printer), pattern arrays can be generated with multiple biopolymer materials.

The present claimed invention further contemplates the optimization of the biopolymer material to maximize the response to given analytes. Although it is not necessary to understand the. mechanism for the purpose of employing the present invention, and it is not intended that the present invention be thereby limited, it has been contemplated that the chemistry of the particular lipid used in the biopolymeric material, plays a critical role in increasing or decreasing the sensitivity of the colorimetric transition. For example, a positional variation of the chromophoric polymer structure can alter the sensitivity to a given analyte. This can be done by moving the diacetylene group closest to the interfacial region as illustrated in Fig. 4 which shows 5, 7-pentacosadiinoic acid (as opposed to 10, 12-pentacosadiinoic acid). In addition, shorter or longer PDA chain lengths were shown to have an effect on analyte permeation due to changes in packaging. In some embodiments, a diacetylene containing from 8 to 28 carbon atoms was used, although shorter and longer chain lengths are contemplated by the present invention. In other embodiments, the position of the diacetylene group may be on the scale of 3 to 16 carbon atoms in the opposite direction from either end of the molecule, although other places are also contemplated by the present invention. Additionally, it has been contemplated that prepared materials of sol-gel of various thicknesses will possess sensitivities to the singular analytes. Thicker films have a higher surface to volume ratio and therefore will require a higher analyte concentration to trigger the chromatic transition. In certain cases, it may be desirable to have a preceptor that is less sensitive. This will prevent the "false trigger" in the presence of low levels of analytes where those levels are not relevant (eg, VOC safety levels), Consequently, the preceptor will find fine tuning to trigger only at or above pre-designated analyte levels. In addition, the gelation conditions of the sol-gel preparation can be optimized by varying gelation temperatures, gel materials and drying conditions to generate material with desired pore dimensions. The variation of the crosslink density of the material also provides control over the pore dimension. The pore dimensions of nanometers to hundreds of nanometers or larger have been contemplated by the present invention. Some gels allow a selective dimension by screening undesirable material, while maintaining the analyte's access to the ligand. Also, the sol-gel technique allows the formation of structures that can be molded into any desired shape including, but not limited to, cartridges, coatings, monoliths, powders and fibers. The sensitivity can also be improved by coupling the lipid-polymer to a photoelectric device, colorimeter or fiber optic tip, which can be read at two or more specific wavelengths. Also, the device can be linked to an alternative signaling device, such as a sound or vibration alarm in order to provide a simple interpretation of the signal.

DESCRIPTION OF THE PREFERRED EMBODIMENTS.

They will be described in the following, specific applications of the claimed invention herein, in order to illustrate the wide application of the invention to a scale of analyte detection systems and to demonstrate its specific application and ease of use.

These examples are intended to illustrate only the broad application of the present invention. The present invention is not intended to be limited to these particular embodiments.

A. Detection of influenza virus. To impart specifications, biological ligand molecules were incorporated into the matrix lipid. For example, a lipophilic sialic acid derivative of carbohydrate was used to specifically bind influenza virus. Sialic acid also has the ability to detect other analytes including, but not limited to, HIV, influnces, chlamydia, Streptococcus suis reovirus, Salmonella, Sendai virus, mumps, newcastle, myxovirus and Neisseria meningi tidis. The invention claimed herein provides superior means for detecting in fl uence, in comparison with the commonly available technology. Immunological tests are limited because of the displacement and movement to antigenic drift exhibited by the virus. The invention claimed herein, detects all varieties of influenza and therefore, a determination of a patient's exposure to influenza will be final and will not be limited to a particular strain. Actually, still uncharacterized newly developed influenza strains can be detected. In some embodiments of the invention claimed herein, polydiacetylene liposome materials coated with sialic acid in a silicate matrix were successfully immobilized by means of the sol-gel method, to provide detector materials that offered optical clarity, and were robust and easy to handle The mild processing conditions allow a quantitative trapping of preformed liposomes without modification of the structure of aggregates. The lipid extraction studies of immobilized unpolymerized liposomes showed no leakage of lipid in aqueous solution for a period of three months. The liposomes of 'polydiacetylene coated with trapped sialic acid, responded to colorimetric signaling to influenza virus X31. Successful transmission of the virus (diameter of 50 to 100 nM) revealed a large pore diameter of the gel that connects the liposome to the volumetric solution. The porous and durable silicate matrix also provides a protective barrier against biological attack (for example, bacteria and fungi) and allows easy recirculation of the liposome preceptor.

The blue to red transition of the polymerized ADCA-containing liposomes was used as a method to detect the presence of virus particles. It was first demonstrated that polymerized liposomes functionalized with sialic acid lipid analogues can bind to influenza virus (Charych et al Science 261: 585 [1993]). Hemagglutinin, the surface lectin of influenza virus, binds to the terminal alpha-glycosides of sialic acid, glycolipids and cell surface glycoproteins (Paulson, The Receptors, Academic Press, New York, Vol. 2, pp. 131 -219 [1985] The purple color of the liposomes containing PDA bound to 5% sialic acid was retained after encapsulation in the sol-gel matrix prepared as monoliths with a thickness of 1-2 mm. shows in the spectrum of Figure 5. However, after a prolonged incubation with influenza virus X31, the color gradually changed towards the red phase as shown in the spectrum of figure 6. The color change was slower , compared to the corresponding solution experiment (ie, the experiment with non-immobilized liposomes in sol-gel glass) .The large spherical dimension of these negatively charged virus particles (50-100 nm in diameter), can inhi Its diffusion through the porous gel is significantly improved, as illustrated in FIG. 7. However, it is not necessary to understand this mechanism in order to use the present invention. However, its ability to reach the liposomes provides some knowledge in the maximum dimension of the gel pores. The attack of bacteria and fungi on the liposomes, observed in solution and with entrapment on agar within the days of preparation (e.g., development of the colony), was not observed for several months encapsulated in accordance with the present invention. The pore size of the gel likewise provides a level of selectivity in these detector materials, separating by sieving the larger interferers, such as bacteria cells (micron dimension), while allowing the selective permeation of smaller agents, such as virus (tens of nanometers). It is contemplated that by altering the gel conditions, the pore dimensions can be controlled to allow for the optimal interaction of a given ligand with a variety of differently sized analytes with controllable response times. In other embodiments, the polymerization of the biopolymer material was conducted. for variable lengths of time to produce different color patterns. Ultraviolet irradiation of PCA liposomes bonded with sialic acid for about 5-10 minutes, results in the formation of an intense blue color as described above. Irradiation for 10-30 minutes produced a solution with a purple color. When the influenza virus was added to the liposomes, a pink or orange color was developed from the blue and purple solutions, respectively. It has also been contemplated that the influenza virus detection system includes additional ligands that recognize and differentiate influenza strains or serotypes from one another and from other pathogenic organisms.

B. Detection of other pathogenic organisms The present invention can also be used to detect a variety of other pathogenic organisms. Ligands, specific for a large number of pathogenic organisms (e.g., carbohydrates, proteins and antibodies) can be incorporated into the biopolymeric material, using routine chemical synthesis methods known in the art (e.g., the globule industry of latex, has shown synthesis procedures for the union of large varieties of chemical groups on synthetic materials). Some of the examples of pathogen detection systems are shown below to demonstrate the variety of methods that can be applied using the present invention and to demonstrate the broad detection capability of single ligand species (e.g., sialic acid). The sialic acid derivative material of the present invention has been used to detect the presence of parasites such as Plasmodium (ie, the etiological agent that causes malaria). In these embodiments, the genetically conserved host binding site was used. PDA films containing sialic acid, as described above, were exposed to solutions containing malaria parasites and erythrocytes. After exposure to parasites overnight, the films turned pink. The color response (CR) in each case was close to 100%. It has been contemplated that the system can be used in conjunction with other test material (e.g., arrays of biopolymer material with several ligands) to identify and differentiate the presence of particularly virulent Plasmodium strains or species. (for example, P. falciparum) or other pathogenic organisms.

In another embodiment, a ligand (ie, GMI ganglioside) was directly incorporated into the biopolymer matrix as described in Example 3 (ie, it was not covalently bound to the diacetylene matrix material). Biopolymeric liposomes of that material colorimetrically detected the presence of cholera toxin, as shown in Figure 12. In this figure, A) and B) show visible absorption spectra of the material before and after exposure to cholera toxin respectively. In still other embodiments, antibodies were used as ligands to detect the presence of Neisseria gonorrhoeae and Vibrio vulnificus. Incorporation of the antibodies into the biopolymeric material has been described in example 3. As is clear from these examples, the present invention provides a variety of means to detect a wide range of pathogenic organisms, including bacteria, viruses and parasites. .

Detection of volatile organic compounds.

Certain embodiments of the invention claimed herein provide means to colorimetrically detect volatile organic compounds (VOC). Most common methods of VOC detection require samples to be taken from laboratory facilities, where they are analyzed by gas chromatography / mass spectroscopy. Some of the methodologies on the site require large and bulky pieces of equipment such as those used in the spectroscopic analysis. Although these methods are excellent, to provide quantitative and contaminant identification, they can not ensure the safety of the individual worker. In one embodiment, the present invention provides a rosette containing trapped biopolymer material that signals the presence of harmful VOCs and provides maximum template security within areas containing VOC. The identity plate is easy and simple to read and does not require experience and analysis by the user. The change of color of the identity plate, indicates to the individual the appropriate action to take. Identity plates reduce costs and improve the efficiency of environmental management and restoration actions, significantly reducing the time due to worker illness preventing exposure to potentially dangerous substances. Two main solutions have been adopted with respect to VOC detection by several groups. The first comprises traditional analytical techniques, such as GC / MS that have been modified for the detection of VOCs (ie, a resolution based on instruments) (Karpe et al., J. Chromatography A 708: 105 [1995]). However, these methods are expensive, complicated and do not lead to their use in the field or at home. The second comprises the coupling of lipid membranes to surface or detector surfaces (that is, a solution with an organic device). In the past decade, several sensing devices have been investigated that comprise the coating of a piezoelectric mass balanced with an organic film. Because of the non-selective nature of the coating, they have been investigated in an array. These sensors, such as quartz crystal microbalance (MCQ) and surface acoustic wave (OAS) devices (see for example Rose-Pehrsson et al., Anal. Chem. 60: 2801 [1988]) have frequency changes. linear with applied masses. By applying a polymer. or another coating on the glass, a sensor based on MCQ or OAS is built. The complex electronics involved in the use of OAS and QCM and electrode-based systems make these solutions less reliable for use as secure personal devices.

The present invention differs from these methods in that the signal transduction is an integral part of the organic layer structure instead of the signal transduction to an electronic device, in addition, embodiments of the present invention, facilitate the optical detection of the signal instead of the electronic detection. In addition, the present invention provides flexibility in the design of material allowing for easy immobilization in a small cartridge (e.g., an identity plate) instead of being etched with the need for electronic equipment. In some embodiments of the present invention, surfactant lipid molecules are used. The use of surfactant liquid molecules on substratum devices allows a standard response to a given VOC depending on the chemical nature of the lipid molecule and the particular VOC. In these embodiments, the advantages of using surfactant molecules in order to build the device through molecular self-assembly are exploited. In aqueous solution, the surfactant molecules are assembled spontaneously in micelles, ampoules, two-layer sheets or thin films, by means of a process that is entropically activated. In addition, materials trapped in sol-gel add robustness and stability. The present invention offers the added benefit of imparting color to the lipid device. During the development of the present invention, it was observed that the interaction of volatile solvents with certain lipid-polymer membranes can produce a transition from strong blue to red. Figure 8, curve a, shows the spectrum of "absorption of a p-PCA film in blue phase." The film changes to the red phase of p-PCA, curve b, under exposure to approximately 500 ppm of 1-octane dissolved in For a variety of solvents analyzed, the degree of color change was generally dependent on the concentration of the solvent and also increased with the degree of halogenation and aromaticity.In this study, a single thin membrane film was prepared. component of p-PCA and polymerized to blue state by UV exposure (254 nm) .These materials were more sensitive to water-immiscible solvents than to water-miscible solvents.For miscible alcohols, the response was found to increase dramatically for isopropanol compared to ethanol, perhaps due to a much higher degree of solvent intercalation in the membrane For the water-immiscible solvents, the measurable changes Color was obtained at 0.05% by weight (500 ppm). Within this group, a similar trend was observed with an increased alcohol chain length, as well as an increased degree of chlorination. A wide variety of water immiscible solvents were examined at their water saturation concentration, as shown in Figure 9. As indicated (Fig 9B), each concentration is different. In Figure 9a, the axis of the "Y" represents the color response or the degree of conversion from blue to red. The numbers above the bar represent an upper limit to the detection in ppm. For many of these solvents, it is clear that solvent concentrations well below 500 ppm can be detected. For the immiscible solvents having a relatively high water solubility, it was possible to examine the effect of solvent concentration on a colorimetric response. It was found that there is a linear relationship between the colorimetric response and the concentration of solvent in water in the range of 0.05-8% by weight, as shown in Figure 10 for 1-butanol. In some embodiments, the claimed invention contemplates the generation of a large palette of polymerizable lipids from different chemistries of front groups to create an array. Lipids containing front groups with carboxylic acid functionalities (imparting a formal negative charge), functionalities of hydrophilic uncharged hydroxyl groups, primary amine (which can acquire a formal positive charge) and hydrophobic groups can be generated. In some embodiments of the present invention, the combination of these materials in a single device facilitates the simultaneous detection of a variety of VOCs or the discrimination of a desired VOC of background materials. In other embodiments, the sol-gel glass material is customary, such that the pores serve as a size-selective sieve to exclude potential interferers from the test material. In addition, the sun processes. The gel of the present invention allows forming structures that can be molded in any desirable manner. For example, in addition to its formulation in a convenient usable cartridge, the coatings can also be prepared on cuvettes and microtiter plates for rapid screening tests. Finally, the lipid-polymer membrane can be coated on other optical measuring device such as a colorimeter or fiber optic tips, used for cases where visual observation is inappropriate or does not provide the required level of sensitivity. Changes in the optical signal must be coupled to an audio or vibration "alarm" signal that provides a secondary warning level. Sol-gel materials can also be processed to form monoliths, powders and fibers. This variability in the configuration or shape of the materials allows the application of the preceptive materials based on polymeric material of the present invention, to most of the platforms, thereby improving the portable condition, handling, durability, sensitivity and Storage time. The pharmaceutical industry has a need that goes to solvent detectors, since pharmaceutical compounds are typically manufactured by means of organic chemical reactions that are carried out in the presence of solvents. Before the packaging of a drug for human use or in other animals, the solvent must be completely removed (Carey and Kowalski, Anal, Chem. 60: 541 [1988]). The method commonly used to detect these VOCs uses intense energy dryers to blow hot air through the drug and piezoelectric crystal arrays to analyze the evaporation of the various solvents (Carey, Trends in Anal. Chem. 13: 210 [1993] ). The invention claimed herein provides a solution based on colorimetry that greatly simplifies these measurements. In addition, the interest in analytical methods for VOC quantifications in non-industrial indoor air environments has increased dramatically in the last 5 years. This is mainly due to a high awareness of emissions of common household appliances or office equipment, as well as trends to control the ventilation of buildings. Companies that produce products for consumers are interested in serving this increased need, providing indoor air monitors that can deduce the presence of hazardous VOCs in you, without the need to sample the air and subsequently analyze in the laboratory. The invention claimed herein, provides embodiments for achieving those means. Indeed, embodiments of the present invention provide improved air sampling and cartridges can be connected to small, portable pumps operated with batteries for personal or general air sampling D Others and emplos.

The above examples demonstrate the wide scale of analytes detectable by the claimed invention herein, ranging from complex biological organisms (eg, viruses, bacteria and parasites) to simple and small organic molecules (eg, alcohols). A number of other analytes have been successfully detected using ligands linked to biopolymeric material include, without limitation, botulinum neurotoxin detected with ganglioside incorporated in p-PDA (Pan and Charych, Langmuir 13: 1367 [1997]). Therefore, it has been contemplated that numerous types of ligands will be linked to self-assembling monomers using common chemical synthesis techniques known in the art, to detect a broad scale of analytes. Additionally, numerous other types of ligands may be incorporated into the biopolymer matrix without covalent binding to self-assembling monomer. These materials allow the detection of small molecules, pathogenic organisms, bacteria, membrane receptors, membrane fragments, volatile organic compounds (VOCs), enzymes, drugs and many other relevant materials. The claimed invention also finds use as a preceptor in a variety of other applications. The color transition of p-PDA materials is affected by changes in temperature and pH. Therefore, the methods and compositions of the invention claimed herein find use as temperature and pH detectors. Ligands can also be used in the present invention when they function as competitive bonds to the analyte. For example, by measuring the colorimetric response to an analyte in the presence of a natural receptor for the analyte, the amount and / or binding affinity of the natural receptor can be determined. Application of competition or inhibition techniques allow the testing of very small non-reactive compounds, as well as substances present in very low concentrations or substances that have a small number or a simple valence. An application of this technique finds use as a means for the development and improvement of drugs by providing a screening test to observe competitive inhibition of natural binding events. The compositions of the invention claimed herein further provide means for testing libraries of materials, since the binding of the desired material can be observed colorimetrically and the relevant biopolymeric material with its relevant ligand separated from the others by external aggregation of a particular polymeric structure (for example, separation of a small portion of sol-gel material contained in an array.

EXPERIMENTAL The following examples are provided in order to demonstrate and further illustrate certain preferred embodiments and aspects of the present invention and have not been construed as limiting the scope of the invention. In the experimental exposition described below, the following abbreviations apply: N (normal); M (molar); mM (millimolar); μM (micromolar); mol (moles); mmol (millimoles); μmol (micromoles); nmol (nanomoles); pmol (picomoles): g (grams); mg (milligrams); μg (micrograms); ng (nanograms); 1 or L (liters); ml (milliliters); μl (microliths); cm (centimeters); mm (millimeters); μm (micrometers); nm (nanometers); μCi (microcurie); mN (milinewton); N (newton); ° C (degrees centigrade); aq (aqueous); J (Joule); PDA (diacetylene monomer); p-PDA (polymerized diacetylene); PCA (pentacosadiinoic acid monomer); p-PCA (polymerized pentacosadiinoic acid); OTS (octadecyltrichlorosilane); VOC (volatile organic chemical), CR (colorimetric response); pH (concentration of hydrogen ions); AFM (atomic force microscope); Hz (Hertz); LB (Langmuir-Blodgett); C02 (carbon dioxide); Sigma (Sigma Chemical Co., St. Louis, Mo); Perkin-Elmer (Perkin-Elmer Co., Norwalk, CT); Fisher (Fisher Scientific, Pittsburg, PA); and Farchan Laboratories (Farchan Laboratories, Inc., Gainesville, FL); Park Scientific Instrument (Park Scientific Instruments, Sunnyvale, CA); Biorad (Bio-Rad Laboratories, Hercules, CA); and Bélico Glass (Bélico Glass Inc., Vineland, BJ). All compounds were pure grade reagents and were used as supplied, unless otherwise stated. The organic solvents were Fischer Scientific spectral grade. All aqueous solutions were prepared "from purified water through a pure Barnstead Type D4700NANO analytical deionization system with a non-organic cartridge that records a resistance of 18.0 M-Ohm-cm.

EXAMPLE Preparation of biopolymeric material.

Production of liposomes _ Self-assembling monomers to be incorporated into liposomes, were dissolved in a solvent (for example, chloroform for diacetylenes, methanol for GMI ganglioside) • Many other volatile compounds find use in the present invention, including without limitation, benzene, hexane and ethyl acetate. The solvent solutions were mixed in appropriate volumes to achieve the desired lipid mixture (eg, 5% per mole of GMI, 95% PCA) and a total lipid content of 2 μmol. Then, the solvent was evaporated by rotary evaporation or with a stream of nitrogen gas. The dried lipids were resuspended in sufficient deionized water to produce a 1-15 mM lipid solution. Then the solution was sonicated for 15-60 minutes with a sonicator probe (Fisher sonic Dismembrador model 300, 50% output, microtip) as described by New (New, supra). The solution was heated during sonification (in most cases the sonication process only provides sufficient heat) at a temperature above the phase transition of the lipids used (typically 30-90 ° C). The resulting mixture was filtered through a 0.8 micromolar nylon filter (Gelman) and cooled to 4 ° C for storage or polymerized. In one embodiment, prior to polymerization, oxygen was removed in the solution, bubbling nitrogen through the sample for 5-10 minutes. Polymerization of the stirred liposome solution was conducted in a 1 cm quartz cuvette with a small UV lamp of 254 nm (lightning pen, energy: 1600 microwatt / cm) at a distance of 3 cm. The chamber was purged with nitrogen during the polymerization to replace all the oxygen and cool the mixture. Polymerization times varied between 5 and 30 minutes depending on the desired properties (e.g., color, degree of polymerization) of the liposomes. In another embodiment, the solution was placed in a UV chamber, without purging and exposed to 0.3-20 J / cm of ultraviolet radiation, preferably 1.6 Jcm, for 5-30 minutes.

Film production Polydiacetylene films were formed in a common Langmuir-Blodgett tundish (see, for example, Roberts, Langmuir Blodget Films, Plenum, New York [1990]).

The trough was filled with water to create a surface for the film. Distilled water was purified with a milli pore water purifier with a resistivity of 18.2 M-Ohm. Diacetylene monomers (eg, 5,7-docosadiinoic acid, 10, 12-pentacosadiinoic acid [Farchan Laboratories], 5, 7-pentacosadiinoic acid, combinations thereof or other self-assembling monomers), dissolved in a solvent spreading agent (eg, spectral grade chloroform [Fisher]), layered on the aqueous surface with a syringe , to form a continuous movie.

Monomers prepared in the concentration scale of 1.0 to 2. 5 nM were maintained at a temperature of 4 ° C in the dark and were allowed to equilibrate at room temperature before being used in experiments. Once layered on the surface, the film was physically compressed using mobile barriers to form a tight packed mono layer of the self-assembling monomers. The monolayer was compressed to its tighter packaged shape (ie, until a film surface pressure of 20-40 nN / M was reached). After compression, the film was polymerized. Ultraviolet irradiation was used to polymerize the monomers, although other polymerization media are available (e.g., gamma radiation, X-ray radiation, and electron beam exposure). Pressure was maintained on the film with the mobile barriers throughout the process of irradiction at the surface pressure of 20-40 mN / m. An ultraviolet lamp was placed 20 cm or more away from the film and the tundish. It was found that if the lamp was placed closer to the film, damage to the diacetylene film can occur due to the heating effects of the film. The film was exposed to ultraviolet light with a wavelength of approximately 254 nm for about one minute. Polymerization was confirmed by observing the acquired blue color under the formation of p-PCA and detecting the linear striations typical of p-PCA films with a polarization optical microscope.

EXAMPLE 2 Sun-Gel entrapment A silica sol was prepared by sonicating 15.25 g of tetramethylorthosilicate (TMOS), 3.35 g of water and 0.22 ml of 0.04 N aqueous hydrochloric acid in a cooling bath until the solution was one phase (approximately 20 minutes). The cooled MOPS buffer solution (50% v / v /) was then added to the acid sol, making sure that the solution was well cooled in an ice bath to retard gelatinization. A variety of materials are suitable for generating silica sols, including, without limitation, any organically modified tetraalkoxysilane or silane (e.g., ormosil). Additionally, tetraethylorthosilicate (TEOS), methyltriethoxysilane (MeTEOS), aryl silsesquioxanes and other metal oxides find use to generate sol-gel glass. To encapsulate liposomes, a solution (2.5 mL) of polymerized liposomes (such as those generated in Example 1) was then mixed in the buffered sol (10 mL) and the mixture was emptied into plastic trays, applied as a film on a flat surface, or was emptied into any other desired forming template, sealed with Parafilm and allowed to remain at room temperature. The gelatinization of the samples occurred within a few minutes resulting in transparent monolithic solids (18 mm x 10 mm x 5 mm) in the case of gels formed in buckets and as violet monoliths with p-PDA liposomes. A slight shrinkage of the aged monoliths was observed, due to the syneresis. The encapsulation of other configurations of biopolymer material (i.e., film and other nanostructures) can be conducted as described above. The materials must be generated or sectioned into small portions if it is not already there, and incorporated in a solution to mix with the dampened sun.

EXAMPLE 3 Ligand binding The self-assembling monomers can be synthesized to contain a wide variety of chemical functions of front groups using synthesis techniques common in the art. The ligands are then linked to the self-assembling monomers by chemical reaction with these functions using synthesis methods well known in the art. Functions include, but are not limited to, esters, ethers, amino, amides or combinations thereof. Alternatively, many ligands can be incorporated into the self-assembling matrix without covalent attachment to the surfactants (eg, membrane proteins and molecules with hydrophobic regions such as gangliosides and lipoproteins). Sialic acid ligand was linked to diacetylene monomers. Various synthesis methods are well known in the art. In one embodiment, PDA (1.0 g, 2.7 mmol in chloroform was reacted with N-hydroxy succinimide (NHS) (0.345 g, 3.0 mmol) and l- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDC) (= : 596 g, 3.1 mmol) The solution was stirred for 2 hours followed by evaporation of the chloroform.The residue was extracted with diethyl ether and water.The organic layer was dried with magnesium sulfate and filtered, then the solvent was evaporated by rotary evaporation to give 1.21 g of N-succinimideyl-PDA (NHS-PDA) Ethanolamine (0.200 mL, 2.9 mmol) was added to a solution of NHS-PDA (1.21 g in 50 mL of chloroform), followed by triethylamine (0.350 mL, 2.5 mmol) and stirred for two hours at room temperature The solvent was evaporated and the residue was purified by silica gel chromatography (2: 1 EtOAc: hexane, Rf = 0.15) to give 0.99 g of N- (2-hydroxyethyl) -APDA. Tetraethyleneglycoldiamine (1.26 g, 6.60 mmol) in 25 mL of chloroform was added to a solution of N- Succinimidyl-PDA (0.603 g, 1.28 mmol) in 20 mL of chloroform, dropwise with stirring for a period of 30 min. The reaction was stirred for an additional 30 min. Before the solvent removal by rotary evaporation. The residue was dissolved in EtOAc and extracted twice with water. The organic layer was dried with magnesium sulfate and the solvent was removed by rotary evaporation. The extract was purified by silica gel chromatography (20: 1 CHCl 3: MeOH, Rf = 0.20) to give 3.72 g of N- (ll-amino-3,6,9-trioxyundecanyl) -APDA. 2 mL of acetic anhydride were added to a cooled solution of ethyl-5-N-acetyl-2,6-anhydro-3,5-dideoxy-2-C- (2-propenyl) -D-erythro-L-hand nononato (0.47 g, 1-30 mmol) in 1.7 ml of pyridine under nitrogen, with stirring. The reaction was allowed to warm to room temperature overnight. After 18 hours, the solvents were removed under reduced pressure at room temperature to produce a crude viscous oil. The oil was solidified by repeated evaporation of toluene. The crude solid was flash chromatographed on silica with ethyl acetate as eluent to yield 0.58 g of ethyl-5-N-acetyl-4,7,8,9-tetra-0-acetyl-3,5-dideoxycarboxylic acid. 2-C- (2-propenyl) -D-erythro-L-mano-nononato. A solution of ethyl-5-N-acetyl-4, 7, 8, 9-tetra-O-acetyl-3,5-dideoxy-2-C- (2-propenyl) -D-erythro-L-mano-nononato (0.38 g, 0.72 mmol) in 10 ml of acetone was cooled to -78 ° C while protecting from moisture with a tube dried with calcium chloride. The ozone was sucked into the solution until the characteristic blue color persisted for 5 mon. The reaction was purged with molecular oxygen to dissipate the excess ozone, followed by heating to 5 ° C. Jones reagent was added in excess (7 drops) until an oxidized orange color persisted and then the reaction was warmed to room temperature. After several minutes, ethanol was added dropwise to consume the excess oxidant. The green precipitate was filtered and washed with acetone several times. The combined filtrates were evaporated in vacuo and dissolved in ethyl acetate. The solution was extracted with a solution of saturated aqueous sodium bicarbonate three times. The combined aqueous layers were acidified with concentrated hydrochloric acid and extracted 5 times with methylene chloride. The combined methylene chloride extracts were dried with magnesium sulfate, filtered and evaporated in vacuo to give ethyl-5-N-acetyl-4,7,8,8-tetra-0-acetyl-3,5-dideoxycarboxylic acid. 2-C- (acetic acid) -D-erythro-L-mannonate. Ethyl-5-N-acetyl-4,7,8,9-tetra-0-acetyl-3,5-dideoxy-2-C- (acetic acid) -D-erythro-L-mannonate 0.43 g. 0.22 mmol and N- (11-amino-3,6,9-trioxyundecanyl) -PDA (0.133 g, 0.24 mmol) were dissolved in 2 ml. of chloroform and the reaction was sealed and stirred for 56 hours. The solution was diluted with 15 ml. of chloroform and the reaction was sealed and stirred for 56 hours. The solution was diluted with 15 ml of chloroform and washed with saturated sodium chloride 1N hydrochloric acid (ag.), Twice; saturated sodium bicarbonate (ag.) 2 times; and saturated sodium chloride (ag.), 1 time. The organic layer was dried over magnesium sulfate, filtered and evaporated to a crude semi-solid. The material was chromatographed by evaporation on silica (20: 1 CHCl 3: MeOH), to yield ethyl-5-N-acetyl-4,5,8,9-tetra-O-acetyl-3,5-dideoxy-2-C - [(N-ll'- (PDA) -3 ', 6', 9'-trioxyundecanyl) -acedamido] -Deritro-L-mano-nononato. PDA derived from sialic acid was formed by dissolving ethyl-5-N-acetyl-4, 5,8,9-tetra-0-acetyl-3,5-dideoxy-2-C- [(N-ll '- (PDA) -3 ', 6', 9 '-trioxiundecanil) -acedamido] -Deritro-L-mano-nononato (0.20 g, 0.19 mmol) in a solution of 4 mL of water and 0.5 mL of methanol containing 0.1 g of hydroxide dissolved sodium. The solution was stirred for 3 hours and an ion exchange resin (Biorad AG 50W-X4H + form) was added until the solution was acidified to a pH paper. The solution was filtered and the filtrate evaporated in vacuo, yielding PDA derived from sialic acid. In other embodiments, carbohydrates (including sialic acid) can be modified by a three step process to produce N-allyl glycosides. The N-allyl glycosides can then be readily linked to other molecules (e.g., PDA) using simple chemical synthesis means routine in the art. This method provides a means to incorporate a broad scale of carbohydrates into biopolymer material (and therefore provides a means to detect a broad scale of analytes). First, the oligosaccharides are dissolved in clean allylamine (if necessary water can be added and the yield is not adversely affected) producing a 0.5 - 0.1 M solution. The reaction is stopped and stirred for at least 48 hours (can be remove small aliquots and assay to complete the reaction as described below). Upon completion of the conversion of the starting material into an amino glycoside product, the solvent is removed by evaporation and the crude solid is treated with toluene and evaporated to dryness several times. Then the solid is cooled in an ice bath and a 60% pyridine solution, 40% acetic anhydride is added to give a solution containing 500 mole percent excess acetic anhydride. The reaction is protected from moisture, stirred and allowed to warm to room temperature overnight. The solvents are removed by evaporation and the residue is dissolved in toluene and dried by evaporation several times. The crude product is purified by flash chromatography, yielding the form of the three sugars of peracetylated Nac-allyl glycolide. The peracetylated Nac-allyl glycosides are then dissolved in methanolanhydrate to give a 0.1-0.001M solution. Several drops of 1 N NaOMe in MeOH are added and the reaction is stirred at room temperature for three hours. Sufficient Dowex 50 resin (H + form) is added to neutralize the base, then the solution is filtered and evaporated to dryness (it can be conducted if purification is desired by recrystallization) The products are the N-allyl glycosamide form of the hydrates of These synthetic reactions have produced the N-allyl glycosamide forms of a variety of carbohydrates, including, but not limited to, glucose, Nac-glucosamine, fucose, lactose, tri-Nac-cytotriose, Lewisx sulfo analog and Lewisx sialyl analogue GMI ganglioside shows an example of incorporation of a ligand without covalent binding to the self-assembling monomers GMI ganglioside was introduced into the biopolymeric material by combining a solution of GMI ganglioside (sigma) dissolved in methanol with dissolved p-PDA in chloroform and dried The ganglioside contains a hydrophobic region that facilitates its incorporation into self-assembling surfactant structures. the dried solutions were re-suspended in deionized water, the resulting structures contain a mixture of ganglioside and p-PDA. The generation of ligands linked to PDA containing a variety of different frontal group chemical species has been described in example 5, for the detection of VOC. These examples demonstrate the PDA derivation with a wide range of chemical front groups, such as hydrophilic uncharged hydroxyl groups, primary amine functions, amino acid derivatives and hydrophobic groups. These and other modifications are generated by synthesis methods common to the art.

The NHS-PDA, as previously generated, and PDA bound to linden provide functional groups for the binding of proteins and antibodies. The NHS or linked thiol monomers are incorporated into the desired aggregate and polymerized. The NHS or thiol functional groups then provide a surface reaction site for covalent attachment to proteins and antibodies using chemical synthesis reactions common in the art. In another embodiment, a hydrazide functional group can be placed on PDA, allowing binding to aldehyde and ketone groups of proteins and antibodies. These embodiments provide a means to incorporate an extremely broad array of proteins and antibodies onto the biopolymer material.

In other embodiments, several other ligands bound to surfactant can be prepared using condensation reactions that involve an activated carboxylic acid group and a hydroxy or amino nucleophilic group. PDA can be activated with trimethylacetyl chloride under anhydrous conditions to form an active asymmetric anhydride. The anhydride can be treated with excess ethylenediamine or ethanolamine to form ethylenediamine-PDA (EDA-PDA) or ethanolamine-PDA (EA-PDA), respectively. One and a half molar equivalents of triethylamine were added as a catalytic base and the reactions were allowed to proceed for three hours at. room temperature. EDA-PDA and EA-PDA are chromatographically purified using a silica gel column and a chloroform / methanol gradient. The EDA-PDA or EA-PDA are then condensed with carboxylic acid containing ligands (chemically activated as before) to form the polymerizable surfactants bound by ligand. Representative examples of ligands that can be prepared by this method include, without limitation, carbohydrates, nucleotides and biotin. The technique contains numerous other examples of successful linkages or ligand association. The self-assembly monomers may be modified chain lengths or may consist of double or multiple chains. These various combinations of ligands and monomers provide an extremely broad array of biopolymer materials suitable for the detection of a broad scale of analytes, with the desired colorimetric response selectivity and sensitivity.

EXAMPLE 4 Characteristic methods I Visible absorption spectroscope Visible absorption studies were performed using a Hewlett Packard 8452a diode array spectrophotometer. For p-PDA material (ie, films, liposomes and entrapped sol-gel materials), the colorimetric response (CR) was quantified by measuring the percent change in absorption at 626 nm (which imparts the blue color to the material ) relative to the maximum total absorption. In order to quantify the response of a biopolymeric material to a given amount of analyte, the visible absorption spectrum of the biopolymeric material without the analyte was analyzed as Bo = I 626 (l 536 + I ß26) where Bo is defined as the absorption intensity at 625 nm divided by the sum of the absorption intensities at 536 and 626 nm. The biopolymer material exposed to analytes was analyzed in the same way as wherein Ba represents the new ratio of absorbance intensities after incubation with the analyte. The colorimetric response (CR) of a liposome solution is defined as the percentage of change in B under exposure to the analyte CR = [(Bo - Ba) / Bo) X 100Í Data demonstrating these determinations are present in several figures, including figure 9 (which shows colorimetric responses of VOCs), figure 11 (shows the colorimetric response to the influenza virus with A) the blue liposome solution before (solid line) and then (interrupted line) of the viral exposure and B) purple liposome solution before (solid line) and after (interrupted line) of the viral exposure), Figure 12 (shows the colorimetric response of cholera A toxin A) before and B) after the exposure) and 1 figure 6 (shows absorbances of the interactions of the influenza virus with p-PDA entrmpado in sol-gel). In figure 6, a p-PDA / sol-gel liposome monolith was incubated in 50 nM Tris buffered to a pH of 7.0 in a plastic cuvette. An aliquot of 50 μl of influenza A X31 was added to the cuvette and the visible absorption spectra were recorded as a function of time from 360 to 800 nm.

Atomic force microscope A biopolymeric material bound with sialic acid was generated as described in Examples 1-3. The materials, either in sol-gel or alone, were exposed to the influenza virus and the colorometric information was observed visually or with spectroscopy, as described in example 4, and shown in figures 5 and 6 for material of blue and red phase, respectively. For liposomes, a mixture of 1-10% PCA bound to sialic acid was incorporated, as indicated in the previous study, optimal viral binding occurs for mixtures of 1-10% in liposomes (Spevak et al., J Am. Chem. Soc. 161: 1146 [1993]). For liposomes entrapped in silicate glass, 5,7-DCDA was found to provide a more vivid colorimetric response than 10, 12-p-PCA. It is believed that the improved response with 5,7-DCDA was related to the size restrictions of the sol-gel material and the topochemical nature of the conformational changes responsible for the chromatic transitions, although an understanding of the mechanism is not required for the practice of the present invention. In one experiment, irradiation of liposome solutions containing p-PCA bound to sialic acid for 5-10 minutes, resulted in the formation of liposomes of intense blue color, while polymerization for between 10 and 30 minutes resulted in a purple color When the influenza virus was added to the liposomes, the material changed to a pink or orange color, depending on whether the initial preparation was blue or purple, respectively. These color changes were easily visible with the eye alone. Competitive inhibition experiments were performed to demonstrate the specificity of the ligand-analyte interaction. The experiments were performed as described above, but with a slight excess of an O-methyl-neuramático acid, a known inhibitor for the hemagglutination of the influenza virus. The presence of the inhibitor resulted in a non-detectable color change of the biopolymer material.

Detection of VOCs Biopolymeric material was generated as described in Examples 1 and 3 and can be incorporated into a porous sol-gel material using the sol-gel method as described in Example 2. The sol-gel / biopolymeric material is then molded into a desired shape and used directly for VOC detection. VOCs were introduced to a p-PDA film in the chromatic and blue phase transitions, from the blue phase to the red phase were measured by detecting the visible absorption spectra as described in example 4. The colorimetric response was measured for 1-butane, 1-hexanol, 1-octanol, CH2C12, CHCI3, CC14, cyclohexane, diethyl ether, toluene and benzene with the data presented in Figure 9 (as previously described). The above numerical values each represent the scale in parts per million, which produced the indicated colorimetric responses. It can be generated to create an array, a large palette of polymerizable lipids from different chemistries of frontal groups. Figure 13 illustrates the lipids for incorporation into a sol-gel matrix. These can be categorized into five groups based on their functions of the front groups. Compounds 2.4 and 2.5 contain carboxylic acid functions that impart a formal negative charge. Compounds 2.6 and 2.7 contain a hydroxyl group without a hydrophilic charge. Compounds 2.8 and 2.9 have primary amine functions that can acquire a formal positive charge. The amino acid derivative 2.10 can exist with positive, negative or zwitterionic charge. Compounds 2.11-2.13 have hydrophobic front groups. The synthesis of these lipids begins with PDA (2.4) commercially available. The synthesis of all except 2.10, 2.12 and 2.13 can be carried out by coupling the respective frontal group to PDA using the activated N-hydroxysuccinimidyl ester of PDA (NHS-PDA) as described in example 3. The lipid 2.10 of amino acid, it can be prepared in four stages of PDA as shown in figure 14, using lithium aluminum hydride and by converting the alcohol to the corresponding bromide derivative. The brumide is converted to the protected amino acid by reaction with diethyl N-acetimido malonate in acetonitrile with sodium hydride, followed by deprotection. Fluorinated lipids 2.12 and 2.13 can be prepared by the reaction of pentafluorobenzoyl chloride with amino lipids 2.8 and 2.9. All publications and patents mentioned in the foregoing specification are incorporated herein by reference. . Various modifications and variations of the disclosed method and system of the invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the invention has been described in connection with specific preferred embodiments, it will be understood that the invention as claimed has not been unduly limited to those specific embodiments. Actually, various modifications of the modes described for carrying out the invention, which are obvious to those experts in molecular biology or related fields, are intended to be within the scope of the following clauses.

Claims (34)

R E I V I N D I C A C I O N S

1. - Method for immobilizing biopolymer material, comprising the steps of: a) providing: i) a metal oxide; ii) biopolymer material; iii) an acid; iv) a shock absorber; and v) zoning means; b) zoning the metal oxide and the acid with the zoning means to produce a zoned solution; c) add the buffer to the zoned solution to produce a buffered solution; and d) adding the biopolymer material to the buffered solution to produce an organic / inorganic solution.

2. - Method according to claim 1, further comprising the steps of: e) applying the organic / inorganic solution to a forming support to produce an organic / inorganic solution; and f) gelling the organic / inorganic solution, to produce an organic / inorganic device.

3. - Method according to claim 1, wherein the metal oxide comprises tetramethyl orthosilicate.

4. - Method according to claim 1, wherein the biopolymer material is selected from the group consisting of liposomes, films. multilayers, braided, laminar, helical, tubular and fiber shapes, solvated rods, solvated coils and combinations thereof.

5. The method according to claim 1, wherein the biopolymer material comprises a plurality of self-assembling monomers selected from the group consisting of diacetylenes, acetylenes, alkenes, thiophenes, polythiophenes, imides, acrylamides, methacrylates, vinyl ether, malic anhydride, urethanes, allylamines, aniline siloxanes, pyrroles, vinyl pyridinium 'and combinations thereof.

6. The method according to claim 5, wherein the diacetylenes are selected from a group consisting of 5,7-docosadiinoic acid, 10, 12-pentacosadiynoic acid, 5,7-pentacosadiynoic acid and combinations thereof.

7. - Method according to claim 5, wherein the self-assembling monomers contain front groups selected from the group consisting of carboxylic acid, hydroxyl groups, amine groups, amino acid derivatives and hydrophobic groups.

8. - Method according to claim 1, wherein the polymeric material further comprises a ligand.

9. - The method according to claim 8, wherein the ligand is selected from the group consisting of peptides, carbohydrates, nucleic acids, biotin, chromophoric drugs, antigens, chelation compounds, molecular recognition complexes, ionic groups, polymerizable groups, groups linkers, electron donors, electron acceptors, hydrophobic groups, hydrophilic groups, receptor binding groups, antibodies and combinations thereof.

10. - Method according to claim 1, wherein the acid comprises hydrochloric acid.

11. - Method according to claim 1, wherein the buffer comprises 3- (N-morpholino) propanesulfonic acid.

12. - Method according to claim 1, wherein the zoning is conducted at a temperature of 0 to 20 ° C.

13. - An organic / inorganic device produced according to the method of claim 2.

14. - Composition comprising biopolymer material encapsulated in sol-gel glass.

15. - Composition according to claim 14, wherein the sol-gel glass comprises tetramethyl orthosilicate.

16. Composition according to claim 14, wherein the biopolymer material is selected from the group consisting of liposomes, films, multilayers, braided, laminar, helical, tubular and fiber forms, solvated rods solvated coils and combinations thereof.

17. Composition according to claim 14, wherein the biopolymer material comprises self-assembling monomers selected from the group consisting of diacetylenes, acetylenes, alkenes, thiophenes, polythiophenes, imides, acrylamides, methacrylates, vinyl ether, malic anhydride, urethanes, allylamines, aniline siloxanes, pyrrole, vinyl pyridinium and combinations thereof.

18. - The composition according to claim 17, wherein the diacetylenes are selected from a group consisting of 5, 7 -docosadinoic acid, 10,12-pentacosadiinoic acid, 5,7-pentacosadiinoic acid, and combinations thereof

19. - Composition according to claim 17, wherein the self-assembling monomers contain frontal groups selected from the group consisting of carboxylic acid, hydroxyl groups, amino groups, amino acid derivatives and hydrophobic groups.

20. - Composition according to claim 14, wherein the biopolymer material also contains a ligand.

21. - Composition according to claim 20, wherein the ligand is selected from the group consisting of peptides, carbohydrates, nucleic acids, biotin, chromophoric drugs, antigens, chelation compounds, molecular recognition complexes, ionic groups, polymerizable groups, groups linkers, electron donors, electron acceptors, hydrophobic groups, hydrophilic groups, receptor binding groups, antibodies and combinations thereof.

22. - Method for detecting analytes, comprising: a) providing: i) biopolymer material encapsulated in sol-gel glass; ii) a detection means; and iii) one or more analytes; b) exposing the biopolymeric material encapsulated in sol-gel glass to the analyte to produce a response; and c) detecting the response using the detection means.

23. - Method according to claim 22, wherein the sol-gel glass comprises tetramethyl orthosilicate.

24. Method according to claim 22, wherein the biopolymer material is selected from the group consisting of liposomes, multilayer films, braided, laminar, helical, tubular and fiber-shaped, solvated solvated coils and combinations thereof.

25. Method according to claim 22, wherein the biopolymer material comprises a self-assembling monomer selected from the group consisting of diacetylenes, acetylenes, alkenes, thiophenes, polythiophenes, imides, acrylates, methacrylates, vinyl ether, malic anhydride, urethanes, allylamines. , aniline siloxanes, pyrroles, vinyl pyridinium and combinations thereof.

26. The method according to claim 25, wherein the diacetylenes are selected from a group consisting of 5,7-docosadiinoic acid, 10, 12-pentacosadiynoic acid, 5, 7-pentacosadiynoic acid and combinations thereof.

27. The method according to claim 25, wherein the self-assembling monomers contain frontal groups selected from the group consisting of carboxylic acid, hydroxyl groups, amine groups, amino acid derivatives and hydrophobic groups.

28. - Method according to claim 22, wherein the biopolymer material further comprises a ligand.

29. - The method according to claim 28, wherein the ligand is selected from the group consisting of peptides, carbohydrates, nucleic acids, biotin, chromophoric drugs, antigens, chelation compounds, molecular recognition complexes, ionic groups, polymerizable groups, groups linkers, electron donors, electron acceptors, hydrophobic groups, hydrophilic groups, receptor binding groups, antibodies and combinations thereof.

30. Method according to claim 22, wherein the analyte is selected from the group consisting of small molecules, pathogenic organisms, bacteria, membrane receptors, membrane fragments, enzymes, drugs, antibodies and combinations thereof.

31. - Method according to claim 22, wherein the biopolymer material encapsulated in sol-gel glass comprises an identity plate.

32. - Method according to claim 22, wherein the detection means is selected from the group consisting of visual inspection, spectrometer, optical fiber, quartz oscillators, electrode surfaces and scintillation.

33. - The method according to claim 22, wherein the response is used as a measure of competitive binding to quantize and characterize the presence of natural binding sites.

34. - Method according to claim 22, wherein the biopolymer material encapsulated in sol-gel glass comprises an array. SUMMARY The present invention relates to methods and compositions for the direct detection of analytes using color changes that occur in immobilized biopolymer material in response to selective binding of analytes to their surface. In particular, the present invention provides methods and. compositions related to the encapsulation of biopolymer material in metal oxide glass using the sol-gel method.