FR2921065A1 - METHOD FOR MANUFACTURING (CO) POLYMERS WITH MOLECULAR FOOT (S) BY PHOTOPOLYMERIZATION UNDER EVANESCENT WAVES, (CO) POLYMERS OBTAINED AND THEIR APPLICATIONS - Google Patents

Abstract

The present invention relates to the field of the synthesis of (co) polymers with molecular imprint (s) and relates to a process for their manufacture. It also relates to the application of the substances obtained by said process for various applications and the devices that result therefrom.This process is characterized by a photopolymerization in which one side of a substrate carrying the substances to be reacted by radiation is irradiated or light rays having a wavelength between 150 nm and 1200 nm so as to form evanescent waves to form said molecular imprinted (co) polymer (s) and then rinsed with at least one solvent suitable for removing all or part of the monomer or monomers that have not polymerized and all or part of the target entity or molecules, if necessary, by detaching the final product obtained from said substrate.

Description

DESCRIPTION

The present invention relates to the field of the synthesis of polymers and / or copolymers having one or more types of molecular imprints (PEM) and more particularly a new process for manufacturing such (co) polymers. It also relates to the application of the substances obtained by the implementation of said process for various known applications or not as such. In particular, it relates to the field of the manufacture of chemical sensors in the broad sense, namely devices for retaining at least temporarily a chemical or biological substance to detect, quantify and / or diffuse in time or in space, such as the measurement or detection of a gas (eg measurement of pollution, chemical reagents ...), the diffusion of a perfume (deodorants), the detection and / or release of any active ingredient (drug, herbicide, catalyst ...), human or animal substances (cells, proteins ...), viruses, etc.

The present application therefore relates to the original implementation of micro and nanostructuring media techniques, such as synthetic polymers, especially for the realization and optimization of chemical sensors. Currently available biosensors use biomolecules as specific recognition elements. The biomolecules on which the currently manufactured biosensors are based have a low chemical and physical stability, in particular with organic solvents, acids / bases, high temperatures, etc. The molecules are expensive and difficult to obtain, and difficult to integrate into industrial manufacturing processes. . As for existing PEMs, their practical use is hampered by overly complex and / or expensive implementation. In this context, the use of PEM is required as a solution to these limits but with the problem of their synthesis that the present invention proposes to simplify through a new process. The present invention aims to overcome these disadvantages. It relates to a method for producing a molecular imprinted (co) polymer (s), optionally in the form of a (co) polymer film, essentially comprising the steps of: a ) selecting at least one target entity or molecule to be reversibly attached to said (fingerprint) molecular (s) (PEM) polymer (s) to be manufactured; b) reversibly binding to or on said at least one target entity or molecule one or more units of at least one fixing monomer so as to obtain, on the target entity or molecules, one or a plurality of monomers capable of polymerizing between them directly, and / or indirectly via at least one multifunctional crosslinking monomer, in order to form, after polymerization, a three-dimensional polymeric network constituting said molecular imprinted (co) polymer (s); c) contacting, on the first of two faces of a support substrate, the target entities or molecules, the fixing monomer (s) and, where appropriate, the multifunctional crosslinking monomer (s) with at least one suitable photosensitive system; d) photopolymerizing, under the action of at least one radiation or a light ray and with the aid of said photosensitive system or systems, said one or more monomers fixing together, if necessary by connecting them to one another via said or said multifunctional crosslinking monomers while at least a part of the target entities or molecules remains attached to said binding monomer (s), characterized in that, for carrying out this photopolymerization, the other second face of said substrate, which substrate carries the aforementioned reactive substances by said radiation or light rays with a wavelength between 150 nm and 1200 nm so as to form on said first face and in a volume of restricted depth, evanescent waves capable of photopolymerizing said fixing monomers, and the multifunctional crosslinking monomer (s) optionally present to form said (co) polymer or (co) polymer film with molecular imprint (s), without chemically modifying said one or more target entities or molecules during said photopolymerization, and e) then rinsing the (co) polymer or imprinted (co) polymer film ( s) molecular (s) previously obtained with at least one suitable solvent to remove all or part of the fixing monomer (s), the non-polymerized crosslinking monomer (s) and all or part of the target entity or molecules still present , where appropriate, by detaching the final product obtained from its support substrate. It also relates to a polymer or copolymer molecular imprint (s) obtainable by the implementation of the method according to the invention, characterized in that it is in the form of a film of a thickness of between 0.1 nm and 10 m, preferably in the form of a film of constant or substantially constant thickness, and a polymer or copolymer with a molecular imprint (s) capable of be obtained by the implementation of the method according to the invention, characterized in that it is in the form of a spatially structured film, preferably having patterns whose critical dimension is between 50 nm and 1 mm. The subject of the present invention is also the use of a polymer or copolymer with molecular imprint (s) obtained by the implementation of the process according to the invention for the manufacture of a qualitative and / or quantitative detector of the presence of at least one target entity or molecule in a sample to be analyzed as well as a qualitative and / or quantitative detector of at least one target entity or molecule in a sample to be analyzed comprising at least one imprinted (co) polymer (s) molecular (s) obtained by the implementation of said method. The subject of the present invention is also the use of a polymer or copolymer with molecular imprint (s) obtained by implementing the method according to the invention for the manufacture of a reversible sensor of at least one target entity or molecule as well as a reversible sensor of at least one target entity or molecule comprising at least one (co) polymer with molecular imprint (s) (PEM) obtained by the implementation of the method according to invention. Finally, the subject of the present invention is also the use of a polymer or copolymer with molecular imprint (s) obtained by implementing the method according to the invention for the manufacture of an affinity separation device. for at least one target entity or molecule as well as an affinity separation device for at least one target entity or molecule comprising at least one (co) polymer with a molecular imprint (s) obtained by implementing the method according to the invention. The present invention uses the evanescent wave photopolymerization technique to structure said support to be treated, generally in the form of a thin film in a single main step, providing a new way of manufacturing these impression materials (s). ) molecular (s). The present invention solves the problem of shaping molecular impression (s) (also known as Molecularly Imprinted Polymers - MIP) or molecularly imprinted (co) polymers (PEM) with objective of manufacturing chemical sensors in the above sense whose operation is based on the principle of molecular recognition. The photostructuration of PEM by the use of the optical near field makes it possible to obtain a multi-scale structuring generally necessary for the proper functioning of the sensor. Thus, structuring at the nanoscale ensures the smooth running of molecular recognition mechanisms and photostructuration itself allows to obtain the structure corresponding to the operation of the sensor. This makes it possible to optimize both the selectivity of the recognition and the sensitivity of the response of a device based on such nanostructured PEM material at two scales. The invention will be better understood, thanks to the following description, which refers to preferred embodiments, given by way of non-limiting example, and explained with reference to the appended diagrammatic drawings, in which: FIG. a simplified diagram of the basic experimental setup used for the manufacture of the PEMs according to the invention; Figure 2 shows a simplified diagram of a variant of the prism used in Figure 1; Figure 3 is a schematic diagram of another alternative of the PEM manufacturing method according to the invention using two interfering laser sources; FIG. 4 represents a photograph taken by interferential microscopy imaging of a PEM film obtained by the method according to the invention with a device according to FIG. 1; FIG. 5 shows an enlarged view of FIG. 4, taken at atomic force microscopy of a PEM according to example 1. FIG. 6 shows a three-dimensional (AFM) microstructure obtained in a PEM according to FIG. Example 1 with a device according to Figure 3; FIG. 7 represents a simplified diagram of the basic experimental setup for a manufacture of PEM according to example 3; and, Figure 8 is a schematic representation of the transducer operating principle of an optical interferometric sensor using a PEM according to the invention as a recognition element. The subject of the present invention is a process for producing a molecular imprinted (co) polymer (s), optionally in the form of a (co) polymer film, essentially comprising the steps of: a) selecting at least one target moiety or molecule to be reversibly attached to said molecular imprint (PEM) (co) polymer to be manufactured; b) reversibly binding to or on said at least one target entity or molecule one or more units of at least one fixing monomer so as to obtain, on the target entity or molecules, one or a plurality of monomers capable of polymerizing between them directly, and / or indirectly via at least one multifunctional crosslinking monomer, in order to form, after polymerization, a three-dimensional polymeric network constituting said molecular imprinted (co) polymer (s) ( PEM); c) contacting, on the first of two faces of a support substrate, the target entities or molecules, the fixing monomer (s) and, where appropriate, the multifunctional crosslinking monomer (s) with at least one suitable photosensitive system ; d) photopolymerizing, under the action of at least one radiation or a light ray and with the aid of said one or more photosensitive systems, said one or more monomers fixing together, if necessary by connecting them to each other via said one or more multifunctional crosslinking monomers while at least a part of the target entities or molecules remains attached to said fixing monomer (s), characterized in that for this photopolymerization it is irradiated, another, second face of said substrate, which substrate carries the aforesaid reactive substances by said radiation or light rays of a wavelength between 150 nm and 1200 nm so as to form on said first face and in a volume of restricted depth, evanescent waves capable of photopolymerizing said fixing monomers, and any multifunctional cross-linking monomer (s) possibly present to form said (co) polymer or film of (co) ) polymer with molecular imprint (s) (PEM), without chemically modifying said target entity or molecules during said photopolymerization, and in that e) then rinsing the (co) polymer or film of (co) ) molecularly imprinted polymer (s) (PEM) previously obtained with at least one solvent suitable for removing all or part of the fixing monomer (s), the non-polymerized crosslinking monomer (s) and all or part of the or target entities or molecules still present, where appropriate, by detaching the final product obtained from its support substrate. Obtaining the new PEM material used for the manufacture of said sensors is based on the principle of molecular printing. Molecular impression is a technique well known to those skilled in the art. It basically consists in its fundamental principle of creating complementary images of a molecule in a support, in general a synthetic (co) polymer. The molecule or molecules against which fingerprints must be generated (molecule or target entity within the meaning of the present invention) are brought into contact with a monomer mixture bearing functional groups capable of complexing the molecule or molecules by weak bonds, by binding bonds. coordination or by reversible covalent bonds.

As will be explained in more detail below, it is essential that the nature of the binding of each target molecule with the PEM is reversible so as to be able, under given conditions, to detach or untie said one or more molecules or entity from the materials in question. fingerprint (s) that we want to synthesize and that will then be used to detect, capture and / or release the relevant substances according to the desired application or use. According to a particularly simple and advantageous embodiment, the contacting of step c) is carried out by depositing one or more liquid solutions containing the target entity or molecules, the fixing monomer or monomers, where appropriate, or the multifunctional crosslinking monomers, and the at least one photosensitive system on the substrate. Examples of solutions will be given above.

Advantageously, the contacting of step c) takes place in the presence of at least one porogenic agent capable of forming pores in the (co) polymer or (co) polymer film with molecular imprint (s) (PEM) that it is desired to obtain, preferably by adding a solution of a pore-forming solvent or a solution of a pore-forming agent in a solvent of the reactive substances present. This facilitates access / departure between the PEM and said target molecules or entities. Preferably, the process according to the invention uses at least one porogenic agent conferring on the PEM a porous structure whose pore size is between 0.1 nm and 10 m, and preferably chosen from the group formed by aqueous solvents or non-aqueous, organic or non-organic, preferably toluene, chlorinated solvents and / or perfluorinated solvents.

Said solvents may further contain a coporogenic agent of a polymeric nature or the like. According to another characteristic, the method according to the invention is further characterized in that it uses radiation or light rays in the form of a ray or beam of light rays, preferably a laser beam, which is a wavelength between 150 nm and 750 nm, preferably between 500 nm and 550 nm, and more preferably between 510 nm and 520 nm. According to a first variant, said method uses radiation or light rays illuminating the assembly of said second surface of the substrate. This produces a uniform and uniform product thickness. According to a second variant, said method according to the invention is characterized in that it uses radiation or spatially structured light rays illuminating only one or certain surface portions of said second surface of the substrate, preferably at least one illuminated surface portion substantially in the form of at least one line or at least one rectilinear strip or substantially in the form of at least one disk, at least one point and / or at least one an area of square or substantially square surface.

In this way, a so-called structured product is obtained and almost all the geometries are possible, which makes it possible to respond to various requests concerning the various branches of the industry that use this type of product. Advantageously, the spatial structure of the radiation or light rays permitting the photopolymerization results from the interference pattern obtained by combining at least two coherent evanescent waves created by at least two interfered light rays before reaching said first face of the light. substrate on which are the reactive substances at the end of step c). This has the advantage of allowing the manufacture of structures that optimize the optical transduction of the sensor while increasing the specific surface area of the PEM. In a particularly efficient manner, the process according to the invention is further characterized in that the photopolymerization of step d) is carried out by displacing the radiations or light rays, preferably by a controlled displacement, and more preferably by a controlled displacement. and computer programmable. This allows the process to be completely automated and to reduce manufacturing costs and improve quality and / or guarantee a required level of quality. The method according to the invention is characterized in that it implements at least one target entity or molecule chosen from the group formed by: amino acids, oses, lipids, nucleosides, nucleotides, oligomers and polymers obtained from the preceding entities or molecules, such as, in particular, peptides, proteins, nucleic acids, oligosaccharides and polysaccharides; biologically active entities or molecules, such as medicaments, in particular antibiotics, toxins, enzymes, doping agents, pesticides, insecticides, fungicides and herbicides, - explosive substances, - nanoparticles, - animal or human cells, such as bacteria, yeasts and eukaryotic cells, - viruses, - mushrooms, and cell tissues. Obviously, it is possible in the context of the present invention to manufacture such materials having only one type of imprint (intended to fix only one entity or target molecule, for example a single specific antibody ) as can be provided several different types of fingerprints (for example for a series of molecules of similar nature or size or having the same type of chemical functionality or biological receptor), whatever the density of the imprint or different types of imprints present on the surface (thin film) and or in the material (cage, three-dimensional network), that is to say the number of impressions per m2 or m3. By way of non-limiting example, the density of the imprints may be between 1018 and 1028 for a size imprint of between 0.5 nm and 100 m. Naturally, the density can be varied according to the type of impression if one is in the presence of a material that has several different kinds of impressions. By material, within the meaning of the present invention, it is necessary to understand all types of polymers or copolymers organic or inorganic usually used for this kind of technique and the skilled person will easily choose the (co) polymers known per se which will be adapted to the implementation of the method according to the invention. Each target entity or molecule has at least one reactive site, preferably several sites of different or identical nature, in which the fixing monomers will be reversibly attached during the step b) of synthesis of the PEM. In the same manner, said fixing molecules may all be identical or different in nature, for example in the form of a mixture of several types of monomers each reacted with a corresponding type of reactive site of the molecule or target entity. According to another characteristic, the process according to the invention is therefore characterized in that it uses at least one fixing monomer chosen from the group formed by the compounds having, on the one hand, a first function allowing at least one bond weak and / or coordination and / or covalent reversible between said one or more fixing monomers, and at least one target entity or molecule thus ensuring at least one reversible bond between said molecular impression polymer (s) (PEM) finally obtained (at the end of the process) and said one or more target entities or molecules and, secondly, at least one second polymerizable function with at least one other identical or different fixing monomer itself or reversibly attached to them at minus one identical or different target molecule, said second function being chosen from the group formed by the monomers comprising at least one radically polymerizable function ical, anionic or cationic, preferably at least one (meth) acrylate, or vinyl.

Advantageously, the fixing monomer or monomers have at least one second polymerizable function with another crosslinking monomer chosen from the group formed by the monomers comprising at least one polymerizable function by radical, anionic or cationic polymerization, preferably at least one (meth) function. acrylate, or vinyl. Said fixing compounds have as a second function a function chosen from the group formed by the hydroxyl, acid, ester, amide, amine, thiol, ether, amidine, guanidine, aldehyde and ketone functions.

Thus, in order to create the molecular imprinting material (s) or the PEM itself, each fixing monomer therefore has in addition at least one first functionality or function allowing it to bind reversibly to said target molecule or entity as well. at least one other second functionality or chemical function allowing it to bind, this time permanently (for example, by covalent bonds) to one or more other corresponding functionalities of a fixing monomer of the same type or type different, from the same target molecule or from a fixing monomer of another target and / or at least one suitable intermediate crosslinking monomer.

Preferably, these first reversible functionality (s) and second (s) permanent functionality (s) are, from a steric point of view, sufficiently distant from each other so as to avoid or at least minimize any harmful interaction. By way of nonlimiting example, it is possible, for example, to quote linear or branched chains of the alkyl type having at one of their ends a first reversible binding functionality with a site of the target molecule or entity, for example an -OH group. which will form a weak, reversible binding or hydrogen bridge bond at the site of said target molecule or entity and, at the other end of the chain and / or on laterally branched or branched branches thereof, one or more a plurality of unsaturated groups of the vinyl type capable of forming strong and permanent bonds by polymerization or condensation (covalent bonding) with other fixing or crosslinking monomers or similar reactive species present so as to form a two or three-dimensional structure at least partially surrounding said molecule or target entity. In the case of a three-dimensional network, it will be ensured that the departure of the molecule or target entity, once the PEM has been formed, can still be satisfactorily achieved by generating a sufficiently porous structure with a maximum specific surface area, so that do not block or completely trap target molecules or entities in the formed EMP. Those skilled in the art will be able to adapt the choice of the molecules or monomers that fix and / or crosslink according to the nature, the size and the properties (active site (s)) of the target molecule on a case by case basis. Advantageously, this second function may also be chosen in such a way that the unit already consisting of a target molecule surrounded by fixing monomer (s) permanently affixes to another compatible fixing monomer. another unit of the same type (identical or different target molecule or entity and identical or different monomer reversibly fixed). In this case, little or no crosslinking monomers are needed.

In the case where the presence of one or more crosslinking monomers is desired or indispensable, at least one multifunctional crosslinking monomer chosen from the group consisting of: the monomers polymerizable by radical polymerization, preferably from base of (meth) acrylate or vinyl monomers, by cationic or anionic polymerization, preferably based on epoxides or thiolenes, or - photopolymerizable hybrid materials such as hybrid sol-gel materials prepared from a precursor preferably 3-methacryloxypropyltrimethoxysilane or 3-glycidyloxypropyltrimethoxysilane. As regards the substrate used, the process according to the invention is characterized in that it advantageously uses a homogeneous and isotropic support substrate. Preferably, said support substrate is preferably selected from the group consisting of glass, quartz, silicon, germanium, silicon carbide, tin-indium oxide and all suitable organic or inorganic polymers.

According to another characteristic, the method according to the invention implements at least one photosensitive system preferably having an effective absorption in the spectral range of UV wavelengths between 150 nm and 400 nm or visible between 400 nm and 800 nm. This has the important advantage of being able to work in a wide spectral range encompassing the visible range which makes handling simpler and less expensive. Another advantage of using visible light for polymerization is that it thus becomes possible to work with targets and / or monomers that are not usable (incompatible) with UV radiation. In a particular embodiment, the photosensitive system consists of a single type of compound (s) operating directly by a photoinduced cleavage reaction of the so-called a and / or (3-cleavage type, preferably at least one chosen from ) in the group formed by benzoin ethers, substituted acetophenones, phosphine oxide derivatives, amino-ketones, oxysulfonyl ketones, sulfonyl ketones, metallocenes and more preferably bis (r15-2.4 cyclopentadien- 1-yl) -bis- [2,6-difluoro-3- (1H-pyrrol-1-yl) phenyl] titanium or one of these derivatives and the azo-type compounds, preferentially azobisisobutyronitrile (AIBN). According to one variant, said photosensitive system consists of a combination of two types of compound (s) functioning by hydrogen tearing and / or electron transfer reaction, one photoinitiator and the other, photosensitizer. , said photoinitiator compound (s) being chosen from the group formed by: weakly basic, preferably tertiary, amines, and more preferably hydroxyalkylamines, in particular methyldiethanolamine, benzylamines, aniline derivatives, and more particularly ethyl paradimethylaminobenzoate, N phenylglycine (NPG), and ascorbic acid, said one or more photosensitizing compounds being chosen from the group formed by acridines, preferably Acriflavine or Acridine Orange, phenazines, preferably Safranine O, and oxazines. thiazines, preferably methylene blue or thionine, xanthenes, preferably Eosin Y, Rose Bengal or Erythrosine, rhodamines, thioxanthenes, polymethines, cetocoumarins and thioxanthones, the photosensitive system comprising as active ingredient a combination of Eosin Y and N-phenylglycine (NPG) being particularly preferred. According to another variant, the photosensitive system SP consists of a combination of two types of compound (s) operating by energy transfer, the one photoinitiator and the other photosensitizer, said one or more photoinitiator compounds being preferably chosen from the group formed by benzoin ethers, preferably 2,2-dimethoxy-2-phenyl acetophenone or substituted acetophenones such as 2-hydroxy-2-methyl-1-phenylpropan-1-one or 2- methyl-1- [4- (methylthio) phenyl] -2-morpholinopropan-1-one and said at least one energy transfer-activated photosensitizer compound being selected from the group consisting of thioxanthone derivatives, preferably isopropylthioxanthone; or chlorothioxanthone, couramines or their derivatives. In a particularly interesting way, the method according to the invention is characterized in that, prior to step c), the support is provided on its first face intended to receive the reactive substances, at least one pretreated zone consisting of a layer of a dielectric material, so as to effect a chemical modification of the surface of said substrate to generate co-polymerizable double bonds, chemical grafting or physical adsorption of the photosensitive system and / or the imprinted (co) polymer ( s) molecular (s) (PEM) to manufacture, said dielectric material being preferably deposited and selected from the group consisting of alkoxysilanes. Advantageously, it is provided that, prior to step c), the support is provided on its first face intended to receive the reactive substances, at least one pretreated zone consisting of a layer of a metal able to have properties plasmonic resonance circuit which makes it possible to use the energy of the plasmon wave generated by the interaction between said metal layer and the applied radiation or light radiation, preferably a deposited metal chosen from the group formed by the gold and the money. Said pretreatment forming said pretreated zone is performed on all or all of said first face of the substrate. In a variant, said pretreatment forming said pretreated zone is carried out solely on one or more surface portions of said first surface of said substrate, preferably on at least one surface portion exposed to radiation or light radiation, on at least one line or at least one rectilinear strip on at least one disk, at least one dot and / or at least one square or substantially square surface area. The subject of the present invention is also a polymer or copolymer with molecular fingerprint (s) (PEM) that can be obtained by the implementation of the method according to the invention, characterized in that it is presented under the in the form of a film having a thickness of between 0.1 nm and 10 m, preferably in the form of a film of constant or substantially constant thickness and a polymer or copolymer with molecular imprint (s) (s) ) (PEM) obtainable by the implementation of said method according to the invention, characterized in that it is in the form of a spatially structured film, preferably having patterns whose critical dimension is included between 50 nm and 1 mm. It also relates to the use of a polymer or copolymer with molecular fingerprint (s) (PEM) obtained by the implementation of the method according to the invention for the manufacture of a qualitative and / or quantitative detector the presence of at least one target entity or molecule in a sample to be analyzed as well as a qualitative and / or quantitative detector of at least one target entity or molecule in a sample to be analyzed comprising at least one (co) polymer to molecular fingerprint (s) (PEM) obtained by the implementation of the method according to said invention. Another subject of the present invention is a use of a polymer or copolymer with molecular fingerprint (s) (PEM) obtained by implementing the method according to the invention for the manufacture of a reversible sensor of at least one target entity or molecule as well as the reversible sensor of at least one target entity or molecule comprising at least one (co) polymer with molecular imprint (s) (PEM) obtained by the implementation of said method. Finally, it also relates to the use of a polymer or copolymer with molecular fingerprint (s) (PEM) obtained by the implementation of said method the manufacture of an affinity separation device for at least one target entity or molecule and the affinity separation device for at least one target entity or molecule comprising at least one (co) polymer with molecular imprint (s) (PEM) obtained by the implementation of said method. In a non-limiting example, the target molecule used as a model to demonstrate the impression of the polymer material is propranolol (beta-blocker). The interest of this molecule resides inter alia in its chirality: If one of the enantiomers is used to generate the imprint, the other constitutes an excellent control of the selectivity of the PEM. The selectivity of the crude polymers synthesized for a given molecule can be characterized using various means: use of fluorescent or radiological markers, liquid chromatography (equilibrium tests). In some cases, the practical use of this material for the development of a chemical sensor presupposes the ability to induce structuring on a micrometric scale. To achieve this, the method according to the invention has been developed. It makes it possible to confine the volume of interaction between the light and the material thanks to an irradiation by the evanescent waves. These waves are produced by total reflection of a beam at an interface. The field thus produced has a limited and above all controllable range, which makes it possible to manufacture photopolymer films of thickness comprised in a few nm and a few hundreds of nanometers. It was validated that this concept could be extended to a photopolymerizable mixture compatible with the molecular printing technique. The photosensitive system previously described has been adapted to meet the requirements of the PEM, particularly in terms of acid constant of the various components. The thickness of the film is directly related to the irradiation time and the power of the beam used. Under these conditions, the thickness of the film obtained can be easily fixed between a few nm and a few hundred nm.

Similarly, the lateral dimension of the polymer articles manufactured can easily be reduced to a few hundred grams using a focused beam. In this case, complex structures can be inscribed by moving the laser beam over the prism / formulation interface. Another very interesting point relates to the possibility of creating a porous structure that can be controlled through the use of a pore-forming solvent (for example toluene). By using a spatially non-homogeneous light source, it is possible to induce a structuring of the polymer blocks created by evanescent wave photopolymerization. In particular, it has been demonstrated that the interference pattern created by the combination of two coherent evanescent waves makes it possible to induce a periodic structuring of the printed polymer films, of nanometric thickness. The main fields of application of the invention are biosensors, chemical sensors, etc. The techniques for structuring the surface of possible polymer materials by means of the process according to the invention are of great interest for applications other than sensors. , in particular in displays (optimization of the extraction of light generated in the electroluminescent layers) or in photovoltaic devices (optimization of the interaction of the incident light with the active material) or in coatings for the release of active products. Among the main technical and economic advantages that may be mentioned is that the present invention makes it possible to synthesize PEM either in the form of films of very small thickness or in the form of nanostructures, the polymerization being able to be controlled and confined in a precisely defined space. In situ synthesis on the surface of the transducer is made possible, and allows, a priori a direct use of the sensor. Conventional reaction mixtures known to those skilled in the art can be used by easily rehabilitating them, which opens the way to the broad range of target molecules referred to above. The photopolymerization initiator will be judiciously chosen by those skilled in the art in order not to interfere with the molecular impression. The invention will now be illustrated with the aid of the following non-limiting examples: 1) Example 1:

The table below gives examples of formulations studied in the context of Example 1 of the present invention.

Table 1: Formulations of Example 1 Functions Products Amounts Used Target Molecule S-Propanolol 20 mg Fixing Monomer Methacrylic Acid 52.6 L Crosslinking Monomer Trimethylolpropane 492 pL Trimethacrylate Photosensitive System Eosin Y 2 mg (dye) Photosensitive System N -phenyl glycine 6 mg (initiator) Diglyme 300 L pore-forming solvent Procedure:

A solution A is prepared by weighing 20 mg of S-propanolol and adding 52.6 l of MAA. The whole is sonicated until complete dissolution then the 492 l of trimethylolpropane trimethacrylate are added. A solution B is prepared by weighing 2 mg of Eosin Y and adding 100 μl of diglyme. The whole thing is re-sonicated until complete dissolution.

A solution C is prepared by weighing 6 mg of N-phenyl glycine and adding 200 μl of diglyme. The whole thing is sonicated until complete dissolution. In the dark, solution C is first mixed with solution B. Then solution A is added to the previously obtained solution to obtain the final mixture or solution. As can be seen schematically in FIG. 1, the basic experimental setup for the manufacture of PEM consists essentially of a laser source (for example an argon ionized laser of 5 W wavelength of 514.5 nm from Spectra -Physics) emitting a focused ray, passing through a diaphragm and a lens before being reflected by a mirror to irradiate the final solution to be light cured previously prepared. To do this, the beam first passes through a glass prism with a high refractive index (n = 1.623) which rests on the edges of a kind of bowl or container (of any suitable material, in particular of the aluminum) and which contains said solution whose refractive index is n = 1.48. Said bowl generally has a depth of 1 mm. The aforementioned prism is placed in such a way that a homogeneous (continuous) surface contact is formed between the solution to be light cured and the latter, in particular by avoiding any presence of air bubbles. At this point, evanescent waves are formed at the prism / solution interface, which photopolymerise the above-mentioned solution and form the desired PEM. More precisely, the power of the laser used at said wavelength is 150 mW / cm 2. The laser beam is linearly polarized and has a Gaussian distribution characteristic of a TEMOO type single mode laser. For such a light intensity, the polymerization times range from a few fractions of seconds to a few tens of seconds. The Gaussian raw beam can be used directly to produce the photopolymerization of the mixture, in solution or shaped by the optical device shown schematically in Figure 1. This device then consists of a spatial filter (set consisting of a microscope objective, micro hole and lens convergent) or a rotating frost (set consisting of two lenses and a diffusing blade rotating). In this configuration, the goal is to produce a homogeneous light spot on the surface of the prism. After irradiation, the prism is removed from the cuvette and rinsed with ethanol. This rinsing eliminates the unreacted monomer layer and releases the fingerprints of the target molecules. The evolution of the thickness of the PEM film is proportional (substantially linear) to the logarithm of the light dose used to induce photopolymerization.

The thickness of the PEM film can be easily chosen between 10 nm and 700 nm by adjusting the intensity of the incident wave and the exposure time, that is to say by varying the light dose (received). between about 30 mJ / cm 2 and 1500 mJ / cm 2. According to a variant shown diagrammatically in FIG. 2, the PEM film can be produced on a plane and transparent substrate, and not directly on the prism as described in FIG. 1. In this configuration, and as diagrammatically illustrated in FIG. an index matching liquid is added between the prism (of the type previously used) and a high refractive index flat plate in order to perform the optical coupling between the prism and said "high index" blade. The aim is to eliminate the optical interfaces between these two elements in order to guarantee the production of the evanescent wave only on the interface between said high index plate and the formulation for obtaining the PEM. The use of this configuration makes it possible to form the PEM film on a transparent substrate in the green (approximately 514 nm), the only essential characteristic of which is to have a refractive index greater than that of the PEM formulation used (1.48 in the case described). Figures 4 and 5 illustrate PEM images obtained with the method according to the invention using the device of Figure 1 or a similar device.

For FIG. 4, a film of 500 m × 100 m of molecular imprinting polymer according to the invention can be observed with a thickness of about 40 nm. The formulation in this case is that described in Table 1 of Example 1. The assembly used in this case was that of Figure 1. The applied light dose was 27 mJ / cm 2 (light power 53 mW / cm 2) with an angle of incidence between the light beam and the 70 ° prism normal, thus guaranteeing the formation of an evanescent wave. For Figure 5, there is an image obtained by atomic force microscopy (AFM) which represents a detail of Figure 4 (image size: 1 gm x 1 m). It shows the porous structure obtained by addition in the photopolymerizable formulation of a pore-forming solvent (here diethylene glycol dimethyl ether) in the PEM film obtained thanks to the implementation of the method according to the invention. FIG. 6 represents an example of microstructures obtained in the PEM film manufactured by the process according to the invention for a formulation identical to that of Example 1 but using the device according to FIG. 3 (AFM image). In this case, the lines were obtained by interference between the two coherent evanescent waves. The modulation of thickness is of the order of 100 nm and the period of the structure of the order of 500 nm. The irradiation conditions were as follows: 30 mJ / cm 2 for an incident power in each 53 mW / cm 2 laser beam. According to another variant diagrammatically shown in FIG. 3, it is possible in this configuration to ensure that the beam delivered by the laser or shaped by an optical device (spatial filter or rotating frosted filter) is separated by a suitable optical device (blade Separator or separator cube). The two beams thus formed are then returned inside the prism by a set of mirrors. In the overlap area of the two beams, a classical interference phenomenon occurs. The advantageous consequence is the production of a sinusoidal distribution of the light intensity which will be used to induce a modulation of the surface of the obtained PEM film. The period of this modulation can easily be adjusted between 170 nm and 32 m by modifying the angle between the two aforementioned beams. The structured PEM film can also be formed on a "high-index" blade (see above) by adapting the assembly as described in the example of FIG.

Example 2

In analogy with the approach described in Example 1, a PEM can be synthesized using a so-called azo initiator. The example describes the synthesis of a selective MEP for the herbicide whose active ingredient is 2,4-dichlorophenoxyacetic acid (2,4-D).

Table 2: Formulation of Example 2 Functions Products Amounts Used Target Molecule 2,4-D 1 mmol Fixing Monomer 4-vinylpyridine 4 mmol Crosslinking monomer Trimethylolpropane 8 mmol trimethacrylate Photosensitive system Azo-bis-0.28 mmol (initiator) isobutyronitrile Pore-forming solvent Water / MeOH 4: 1 3 ml The mixture is bubbled with argon and deposited on the high-index glass substrate on which the PEM is to be synthesized. The substrate is then assembled with a prism in an assembly similar or identical to that used in Example 1. The assembly is placed in a chamber under an inert atmosphere and the polymerization is initiated by irradiating the mixture with a laser of a length 364 nm wave. After a suitable polymerization time (of the order of a few seconds), fine spots of PEM are obtained on the surface of the glass substrate. The results obtained are comparable to those of Example 1. Example 3: Polymerization in SPR mode

In analogy with the approach described in Example 1, a PEM can be synthesized in so-called SPR mode (that is Surface Plasmonic Resonance). The example describes the synthesis of a selective PEM for betabloquant propranolol. The formulation used in this example again corresponds to that used in Example 1. On a high-index glass substrate are successively deposited a first layer 2 nm thick Cr as a primer layer advantageously serving to assist the deposition of a second layer of 47 nm thick gold. The two layers may be deposited by any technique known to those skilled in the art, preferably by evaporation. The mixture to be polymerized is bubbled with argon and deposited on the gold-side substrate. The substrate is then assembled with a prism in an assembly such as that used in Example 1. The assembly is also placed in a chamber under an inert atmosphere. The polymerization is initiated by irradiating the reaction mixture with a 532 nm laser, a power of 1 mW, pumped by diode through the prism of Example 1, the liquid solution to be polymerized being deposited on the glass support covered with Cr, itself covered with gold. As can be seen in FIG. 7, the incident laser beam penetrates the trapezoidal vertical section prism (small side down) placed directly under the glass substrate (thickness = 500 m, refractive index = 1.623 ) by the left oblique face, passes through the substrate and is reflected on the gold layer where evanescent waves arise in the solution reacted by photopolymerization, to start symmetrically with respect to the vertical direction (X axis). X ') according to the same angular value (relative to the horizontal axis X-X') and to emerge on the right symmetrical oblique face of said prism.5 After a suitable polymerization time (from a few seconds to a few minutes depending on the desired thickness), fine spots of PEM are obtained on the gold surface.

Example 4: Manufacture of PEM Networks for Application as a Biomimetic Chip

Manufacture: High index glass substrates are silanized by methacryloyl trimethoxysilane allowing the attachment of PEM by copolymerization via covalent bonds. The glass substrate is assembled with a prism for the evanescent wave polymerization as described in Example 1 and the assembly is mounted on an X-Y microdisplacement platen. This makes it possible to move the substrate on which the PEM must be synthesized perpendicularly to the evanescent wave created by the laser beam. After each polymerization step, the substrate is moved in the X and / or Y direction to synthesize a new pad at a defined distance. In this way, a network of PEM pads is finally obtained which constitutes the biomimetic chip. Beside this PEM network, a number of polymer pads without molecular imprints are synthesized as controls.

The chip is washed for 3 times 1 h with a methanol / H 2 O mixture 9: 1, then twice for 30 min. with methanol and dried under nitrogen flow.

Use: Screening of Multiplexed and / or High-Flow Molecules The PEM pads on the chip are contacted with a solution containing a constant concentration of a fluorescent derivative of the target molecule, and another molecule as a competitor. After an incubation time of 1 h, the chip is analyzed by a fluorescence reader to quantify the fluorescence emitted by the PEM pads. If the competitor binds to the recognition sites of the PEM, the fluorescent derivative of the target molecule does not fix or bind to a lesser extent and the corresponding pad (control) is not fluorescent or less fluorescent. The pad made of PEM thus adsorbs a larger number of targets, has a better selectivity and a better affinity than the control pad without imprints. This allows: - to detect the presence of the target molecule in a sample, - to determine the concentration of the target molecule in a sample if a standard curve is previously recorded using a range of concentrations of the target molecule, - to screen , by their cross-reactivity, other molecules for possible affinity for the synthetic receptor (PEM), - to develop high-throughput tests if the pads are addressed individually by a microfluidic system or by localized deposition of the samples to be analyzed, or if all of the pads are addressed by an imaging system using a camera or a scanning fluorescence detection system, - to develop multiplexed tests if several pads of PEM for different targets are synthesized on the chip.

Example 5 Microetching on a Biochip Manufacture of the Chip

High-index glass substrates are silanized by methacryloyl trimethoxysilane allowing attachment of PEM by copolymerization via covalent bonds. The glass substrate is assembled with a prism for the evanescent wave polymerization as described in Example 1, and the assembly is mounted on an X-Y microdisplacement platen. This makes it possible to move the substrate on which the PEM must be synthesized perpendicularly to the evanescent wave created by the laser beam. After each polymerization step, the substrate is moved in the X and / or Y direction to synthesize a new pad at a defined distance. In this way, a network of PEM pads is finally obtained which constitutes the biomimetic chip. Next to this PEM network, a number of polymer pads without molecular imprints are synthesized as controls. The chip is washed for 3 times 1 h with a methanol / H 2 O mixture 9: 1, then twice for 30 min. with methanol and dried under nitrogen flow.

Use :

The sample containing the target molecule is deposited on a pad and left in contact for a sufficiently long time to allow adsorption of the target on the PEM. Then the chip is washed to remove the adsorbed molecules nonspecifically, then an elution solution is deposited on the pad. After incubation, the solution which now contains the target molecule is removed and analyzed by a conventional method for detection and / or quantification of the target. The use of several pads in parallel will make it possible to extract several samples at the same time.

Example 6 Manufacture of PEM Microchannels for Microseparation by Affinity Chromatography

PEMs are synthesized as a rectilinear line or strip on a planar substrate. The substrate is assembled in a microfluidic system that creates a channel along the PEM. At the end of the channel a detector is mounted to detect the passage of the target molecule. With the aid of a pump, a solution containing the target molecule is injected into the channel in a continuous flow. The affinity of the target for the EMP is either retained or slowed by the EMP. In the case of retention, it will be eluted by injection of an elution solvent and collected in a fraction. In the case where it is slowed, fractions are collected at the outlet of the channel and the fraction containing the target molecule 30 identified via the detector signal.

Example 7: Interferometric sensor

The PRM is synthesized according to a method characterized in that the spatial structure of the light ray for photopolymerization results from the interference pattern obtained by combining at least two coherent evanescent waves created by at least two interfering light rays. before reaching the substrate on which the reactive substances are found (see Fig. 3 and 6 with a solution according to Example 1). The microstructured PEM is used in a sensor based on optical interferometry. FIG. 8 describes, as an indication, the general schematic principle of transduction of an optical interferometer sensor using a PEM as a recognition element. The diffraction of the wavelength guided wave is represented before (continuous line) and after (dashed lines) modification of the refractive index of the PEM due to the adsorption of the target molecule.

Of course, the invention is not limited to the embodiments described and shown in the accompanying drawings. Modifications are possible, particularly from the point of view of the constitution of the various elements or by substitution of technical equivalents, without departing from the scope of protection of the invention.

Claims (32)

1) A process for producing a molecular imprinted (co) polymer (s), optionally in the form of a (co) polymer film, essentially comprising the steps of: a) choosing at least one entity or target molecule to be reversibly attached to said molecular imprinted (co) polymer (s) to be made; b) reversibly binding to or on said at least one target entity or molecule one or more units of at least one fixing monomer so as to obtain, on the target entity or molecules, one or a plurality of monomers capable of polymerizing between them directly, and / or indirectly via at least one multifunctional crosslinking monomer, in order to form, after polymerization, a three-dimensional polymeric network constituting said molecular imprinted (co) polymer (s); C) contacting, on the first of two faces of a support substrate, the target entities or molecules, the fixing monomer (s) and, where appropriate, the multifunctional crosslinking monomer (s) with at least one suitable photosensitive system ; d) photopolymerising, under the action of at least one radiation or a light beam and with the aid of said one or more photosensitive systems, said one or more monomers fixing together, if necessary by connecting them to each other via said one or more multifunctional crosslinking monomers while at least a part of the target entities or molecules remains attached to said fixing monomer (s), characterized in that for carrying out this photopolymerization the another, second face of said substrate, which substrate carries the aforesaid reactive substances by said radiation or light rays with a wavelength between 150 nm and 1200 nm so as to form on said first face and in a depth volume restricted, evanescent waves capable of photopolymerizing said fixing monomers, and the multifunctional crosslinking monomer (s) optionally present to form said (co) polymer or film of (co) polymer with molecular imprint (s), without chemically modifying said one or more target entities or molecules during said photopolymerization, and in that e) then rinsing the (co) polymer or film of (co) Molecular impression polymer (s) previously obtained with at least one suitable solvent for removing all or part of the fixing monomer (s), non-polymerized crosslinking monomer (s) and all or part of the entity (s) or molecule (s) or molecule (s) still present targets, if any, by detaching the final product obtained from its support substrate. 2. Method according to claim 1, characterized in that the contacting of step c) is carried out by depositing one or more liquid solutions containing the target entity or molecules or molecules, the fixing monomer or monomers, where appropriate. the multifunctional crosslinking monomer or monomers, and the at least one photosensitive system on the substrate. 3. Method according to claim 1 or 2, characterized in that the contacting of step c) is in the presence of at least one porogenic agent capable of forming pores in the (co) polymer or film of (c) ) Molecular impression polymer (s) that is desired to obtain, preferably by adding a solution of a pore-forming solvent or a solution of a pore-forming agent in a solvent of the reactive substances present. 4. Process according to claim 3, characterized in that it uses at least one porogenic agent conferring on the (co) polymer or (co) polymer film with the obtained molecular number (s) a porous structure whose pore size is between 0.1 nm and 10 m, and preferably selected from the group consisting of aqueous or non-aqueous solvents, organic or otherwise, preferably diethylene glycol dimethyl ether, toluene, chlorinated solvents and / or perfluorinated solvents. 5. Method according to any one of the preceding claims, characterized in that it uses radiation or light rays in the form of a ray or beam of light rays, preferably a laser beam, a wavelength between 150 nm and 750 nm, preferably between 500 nm and 550 nm, and more preferably between 510 nm and 520 nm. 6. Method according to any one of the preceding claims, characterized in that it uses radiation or light rays illuminating all of said second surface of the substrate. 7. Method according to any one of claims 1 to 5, characterized in that it implements spatially structured radiation or light rays illuminating only one or more surface portions of said second surface of the substrate, preferably at least one illuminated surface portion substantially in the form of at least one line or at least one rectilinear strip or substantially in the form of at least one disk, at least one dot and / or at least one area of square or substantially square area. 8. Method according to any one of claims 6 or 7, characterized in that the spatial structure of radiation or light rays for photopolymerization results from the interference pattern obtained by combining at least two coherent evanescent waves created by at least two light rays interfered before reaching said first face of the substrate on which the reactive substances are at the end of step c). 9. Process according to any one of the preceding claims, characterized in that the photopolymerization of step d) is carried out by displacing the radiations or light rays, preferably by a controlled displacement, and more preferably by a controlled displacement and computer programmable. 10. Method according to any one of the preceding claims, characterized in that it implements at least one entity or target molecule selected from the group consisting of: - amino acids, oses, lipids, nucleosides, nucleotides oligomers and polymers obtained from the preceding entities or molecules, such as, in particular, peptides, proteins, nucleic acids, oligosaccharides and polysaccharides; biologically active entities or molecules, such as medicaments, in particular antibiotics, toxins, enzymes, doping agents, pesticides, insecticides, fungicides and herbicides, - explosive substances, - nanoparticles, - animal or human cells, such as bacteria, yeasts and eukaryotic cells, - viruses, - mushrooms, and - cellular tissues. 11. Process according to any one of the preceding claims, characterized in that it uses at least one fixing monomer chosen from the group formed by the compounds having, on the one hand, at least one first polymerizable function with another fixing monomer and / or a crosslinking monomer chosen from the group formed by the monomers comprising at least one radical polymerizable, anionic or cationic polymerizable function, preferably at least one (meth) acrylate or vinyl function. 12. Process according to claim 11, characterized in that it uses at least one fixing monomer chosen from the group formed by the compounds having, on the other hand, a second function allowing at least one weak bond and / or coordination and / or reversible covalent between said one or more fixing monomers, said one or more target entities or molecules thereby ensuring reversible binding between said molecular imprint polymer (s) and said target entity or molecule. 13. The method of claim 12, characterized in that said fixing compounds have as a second function a function selected from the group consisting of hydroxyl functions, acid, ester, amide, amine, thiol, ether, amidine, guanidine, aldehyde and ketone. . 14. Method according to any one of the preceding claims, characterized in that it uses at least one multifunctional crosslinking monomer selected from the group consisting of: - monomers polymerizable by radical polymerization, preferably based on monomers (meth ) acrylate or vinyl, by cationic or anionic polymerization, preferably based on epoxides or thiolenes, or - photopolymerizable hybrid materials such as hybrid sol-gel materials prepared from a precursor, preferably 3-methacryloxypropyl trimethoxysilane or 3-glycidyloxypropyltrimethoxysilane. 15. Method according to any one of the preceding claims, characterized in that it implements a homogeneous and isotropic support substrate. 16. Process according to any one of the preceding claims, characterized in that the support substrate is preferably chosen from the group formed by glass, quartz, silicon, germanium, silicon carbide, tin-indium and all suitable organic or inorganic polymers. 17. Method according to any one of the preceding claims, characterized in that it implements at least one photosensitive system preferably having an effective absorption in the spectral range of UV wavelengths between 150 nm and 400 nm or visible. between 400 nm and 800 nm. 18. The method of claim 17, characterized in that the photosensitive system consists of a single type of compound (s) operating directly by a photoinduced cleavage reaction of the type called a and / or [3-cleavage, preferably at least one selected from the group consisting of benzoin ethers, substituted acetophenones, phosphine oxide derivatives, amino ketones, oxysulfonyl ketones, sulphonyl ketones, metallocenes and more preferably bis ( 15-2,4-cyclopentadien-1-yl) -bis- [2,6-difluoro-3- (1H-pyrrol-1-yl) phenyl] titanium or one of these derivatives and the compounds of the azo type, preferentially azobisisobutyronitrile (AIBN). 19. The method of claim 17, characterized in that the photosensitive system consists of a combination of two types of compound (s) operating by hydrogen tearing reaction and / or electron transfer, a photoaminer and the other, photosensitizer, said photoinitiator compound (s) being chosen from the group formed by: weakly basic, preferably tertiary, amines, and more preferably hydroxyalkylamines, in particular methyldiethanolamine, benzylamines, derivatives of aniline, and more particularly ethyl paradimethylaminobenzoate, N-phenylglycine, and ascorbic acid, said photosensitizing compound (s) being chosen from the group formed by acridines, preferably Acriflavine or Acridine Orange, and phenazines. Safranine O, oxazines, thiazines, preferably methylene blue or thionine, xanthenes, preferably Eosin Y, Rose Bengal or Erythrosine, rhodamines, thioxanthenes, polymethines, cetocoumarins and thioxanthones, the photosensitive system having as active principle a combination of Eosin Y and N-phenylglycine. being particularly preferred. 20. The method of claim 17, characterized in that the photosensitive system consists of a combination of two types of compound (s) operating by energy transfer, the one photoinitiator and the other photosensitizer, said compound or compounds photoinitiators are preferably selected from the group consisting of benzoin ethers, preferably 2,2-dimethoxy-2-phenyl acetophenone or substituted acetophenones such as 2-hydroxy-2-methyl-1-phenylpropan-1 -one or 2-methyl-1- [4- (methylthio) phenyl] -2-morpholinopropan-1-one and said one or more photosensitizer compounds functioning by energy transfer being chosen from the group formed by the thioxanthone derivatives preferably isopropylthioxanthone or chlorothioxanthone, the couramines or their derivatives. 21. Method according to any one of the preceding claims, characterized in that, prior to step c), the support is provided on its first face intended to receive the reactive substances, at least one pretreated area consisting of a layer of a dielectric material, so as to effect a chemical modification of the surface of said substrate to generate co-polymerizable double bonds, chemical grafting or physical adsorption of the photosensitive system and / or imprinted (co) polymer ( s) molecular (s) to manufacture, said dielectric material being preferably deposited and selected from the group consisting of alkoxysilanes. 22. Method according to any one of the preceding claims, characterized in that, prior to step c), the support is provided on its first face intended to receive the reactive substances, at least one pretreated zone consisting of a layer of a metal capable of presenting plasmonic resonance properties making it possible to use the energy of the plasmon wave generated by the interaction between said metal layer and the applied radiation or light radiation, preferably a deposited metal selected in the group formed by gold and silver. 23. The method of claim 21 or 22, characterized in that the pretreatment forming said pretreated zone is performed on all or all of said first face of the substrate. 24. The method of claim 21 or 22, characterized in that the pretreatment forming said pretreated area is performed only on one or more surface portions of said first surface of said substrate, preferably at least on a surface portion exposed to radiation or radiation. at least one line or at least one rectilinear strip on at least one disk, at least one point and / or at least one square or substantially square surface area. Polymer or copolymer with molecular imprint (s) obtainable by implementing the method according to any one of the preceding claims, characterized in that it is in the form of a film of a thickness of between 0.1 nm and 10 m, preferably in the form of a film of constant or substantially constant thickness. 26. Polymer or copolymer with molecular fingerprint (s) obtainable by carrying out the method according to any one of claims 1 to 24 or according to claim 25, characterized in that it is presented in the form of a spatially structured film, preferably having patterns whose critical dimension is between 50 nm and 1 mm. 27. Use of a polymer or copolymer molecular imprint (s) obtained by the implementation of the method according to any one of claims 1 to 24 or claim 25 or 26 for the manufacture of a detector qualitative and / or quantitative of the presence of at least one target entity or molecule in a sample to be analyzed. 28. Qualitative and / or quantitative detector of at least one target entity or molecule in a sample to be analyzed comprising at least one molecular imprinted (co) polymer (s) obtained by implementing the method according to the invention. any of claims 1 to 24 or claim 25 or 26. 29. Use of a polymer or copolymer molecular imprint (s) obtained by the implementation of the process according to any one of claims 1 to 24 or claim 25 or 26 for the manufacture of a sensor reversible of at least one target entity or molecule. 30. Reversible sensor of at least one entity or target molecule comprising at least one molecular imprinted (co) polymer (s) obtained by carrying out the method according to any one of claims 1 to 24. or according to claim 25 or 26. 31. Use of a polymer or copolymer with molecular imprint (s) obtained by the implementation of the process according to any one of claims 1 to 24 or claim 25 or 26 for the manufacture of a device affinity separation for at least one target entity or molecule. An affinity separation device for at least one target entity or molecule comprising at least one molecular imprinted (co) polymer (s) obtained by carrying out the process according to any one of claims 1 to 24. or according to claim 25 or 26.