Method and coating system for immobilising biomolecules

Abstract

The invention relates to methods for producing a coated solid-state substrate for immobilising biomolecules and to coating systems for immobilising biomolecules. The invention also relates to a solid-state substrate for immobilising biomolecules produced by a method according to the invention and comprising the coating system, as well as a functional solution to be used in the method for producing a component of the coating system. Finally, the invention relates to the use of the coating system for immobilising biomolecules as well as a microarray or biochip comprising the coating system for analysing biomolecules contained in a sample.

Description

[0001] 05 / 12 / 2024 Method and coating system for the immobilization of biomolecules Description Method for producing a coated solid substrate for the immobilization of biomolecules as well as coating systems for the immobilization of biomolecules. Furthermore, the invention relates to a solid substrate for the immobilization of biomolecules produced by a method according to the invention and comprising the coating system and a functional solution for producing a component of the coating system. Finally, the invention relates to the use of the coating system for the immobilization of biomolecules as well as a microarray or biochip comprising the coating system for the analysis of biomolecules contained in a sample. Various microarrays and biochips are known in practice that make it possible to immobilize biomolecules contained in a sample in a very small space and present them for further analysis.In this regard, it should be noted that the term "sample" in the context of this disclosure is to be understood in the broadest sense, and thus includes, among other things, body fluids to be analyzed, such as blood, blood plasma, urine, saliva, etc., as well as tissue, tissue sections or whole cells isolated therefrom, but also laboratory-produced samples, such as cell layers, cell extracts, cell culture media and supernatants, nucleic acid or protein solutions, etc., whereby such a sample can be immobilized and ultimately analyzed by immobilizing the biomolecules contained therein on the surface of the array or chip. Reactive coatings for the immobilization of biomolecules, for example on microarrays or biochips, allow a large number of different samples to be applied in a very small space (usually with the help of robots) in defined and delimited areas ("spots"), whereby theThe biomolecules to be analyzed in the respective samples come into contact with the coating and, through this contact, are specifically immobilized in the respective spot for further steps of the analysis process. 05 / 12 / 2024 DNA microarrays (or DNA chips), for example, regularly consist of solid substrates, usually glass plates, comprising a layer of chemically fixed DNA molecules, which in turn interact with complementary DNA molecules of a sample through nucleic acid hybridization, so that the complementary DNA molecules are specifically immobilized in the area of ​​the respective spot. The binding of two complementary DNA molecules can then be optically detected and quantified in the subsequent analysis process, e.g., using fluorescent markers that are coupled to the sample DNA during the analysis. For example, microarrays are used to investigate the expression of a very large number of genes simultaneously or toto diagnose specific gene mutations. Biochips can act as biosensors to represent biologically / medically relevant properties and reactions by (a) comprising a coating on which specific biomolecules of a sample, e.g., nucleic acids, peptides, proteins such as enzymes or antibodies, are immobilized and (b) containing an integrated microprocessor that evaluates the signals of the immobilized biomolecules. Various coatings for the immobilization of biomolecules are also known in practice. Conventional methods for coating solid-state substrates for the production of microarrays and biochips for the analysis of biological samples include any known method for applying adhesion-promoting or functional layers of complex coating systems, including by means of spraying, spraying and / or spin technology. In this context, the use of UV-curable coatings for im-mobilization of biomolecules is also known. A method for producing a reactive, UV-curable coating for immobilizing biomolecules in the context of an enzyme-linked immunosorbent assay (ELISA) is described, for example, in EP 2532639 A1. The described coating for immobilizing peptides is produced using UV-reactive cycloalkanes, in particular bicyclononynes, by means of CLICK chemistry. 05 / 12 / 2024 Another method for producing a reactive, UV-curable coating for immobilizing biomolecules, in particular nucleic acids and proteins, is described in US 2005 / 0074478 A1. To produce the described coating, acrylamide and acrylate polymers are used, which are curable by UV irradiation, so that the coating ultimately comprises acrylamide-acrylate copolymers, whereby the acrylate polymers contain functional groups suitable for the immobilization of biomolecules.UV-curable coatings known from the state of the art for immobilizing biomolecules on coated surfaces, such as the surfaces of microarrays or biochips, are often limited in their binding capacity for biomolecules, so that detection against nonspecific background fluorescence often presents a challenge in practice. Furthermore, conventional UV lamps, typically medium-pressure mercury lamps, are regularly used to cure such coating systems. Since a large portion of the energy consumed by the lamp is not converted into UV radiation, but into visible light and infrared radiation, i.e., heat, conventional lamps must be cooled. There is also a risk of burns from touching the lamp. Furthermore, the use of conventional UV lamps to cure coating systems regularly results in the formation of health-relevant quantities.of the irritant gas ozone. Ozone produced must not escape into the workspace in dangerous concentrations and must be extracted, removed, or absorbed at the point of origin. Therefore, an object of the present invention is to design and further develop a method for producing a solid substrate coated with a coating of the type mentioned above in such a way that a coating system with high binding capacity for biomolecules and low background fluorescence can be produced and provided, as well as to provide a correspondingly coated solid substrate and a functional solution for producing a component of the coating system. A further object of the present invention is to specify both a use of the coating system for immobilizing biomolecules on a surface of a microarray or a biochip provided with the coating system, anda microarray or biochip comprising the coating system, 05 / 12 / 2024, in particular for the analysis of biomolecules contained in a sample. In this regard, it should be noted that the term "sample" in the context of this disclosure is to be understood in the broadest sense and thus includes, among other things, both body fluids to be analyzed, such as blood, blood plasma, urine, saliva, etc., as well as tissue, tissue sections, or whole cells isolated therefrom, but also laboratory-produced samples, such as cell layers, cell extracts, cell culture media and supernatants, nucleic acid or protein solutions, etc., whereby such a sample can be immobilized and ultimately analyzed by immobilizing the biomolecules contained therein on the surface of the array or chip. In one embodiment, the present invention achieves the above-mentioned objects with a method for producing acoated solid substrate for the immobilization of biomolecules comprising the following steps: (a) applying an adhesion promoter layer to at least one surface of the solid substrate, wherein the contact angle of water on the adhesion promoter layer is preferably between 50 ° and 60 °; (b) providing a functional solution comprising: ^ a crosslinking degree modulator comprising: acrylate monomers and / or acrylate oligomers with one acrylate ester group per molecule, preferably tetrahydrofurfuryl methacrylate monomers and / or oligomers; and / or 2-phenoxyethyl acrylate monomers and / or oligomers; and / or acrylate monomers and / or acrylate oligomers with more than one, preferably more than two or more than three acrylate ester groups per molecule, in particular comprising: trimethylolpropane ethoxy triacrylate monomers and / or oligomers; Dipentaerythritol penta- / hexacrylate monomers and / or oligomers; and / or functional dendrimers and / orDendrons, preferably highly branched bis(hydroxymethyl)propionic acid, acetylenic, highly branched bis(hydroxymethyl)propionic acid, azide and / or highly branched bis(hydroxymethyl)propionic acid, hydroxyl and / or highly branched poly(amidoamine) dendrimers and / or dendrons; ^ an immobilization modulator which provides reactive groups for the immobilization of biomolecules, the immobilization modulator preferably comprising: ester groups, preferably N-hydroxysuccinimide ester groups, glycidyl ester groups or isocyanate ester groups; ether groups, preferably glycidyl ether groups; epoxy groups; aldehyde groups; free carboxyl and / or free amino groups, and combinations thereof; and 05 / 12 / 2024 ^ a photoinitiator having an absorption maximum in the wavelength range between 315 and 400 nm, preferably a photoinitiator comprising: phenylglyoxylic acid methyl ester and diphenyl-(2,4,6-trimethylbenzoyl)-phosphine oxide; ethyl-phenyl(2,4,6-trimethylbenzoyl)phosphinate; or phenylbis(2,4,6-trimethylbenzoyl)phosphine oxide; (c) applying the functional solution to the adhesion promoter layer; and (d) curing the functional solution using LED UV-A radiation in the wavelength range between 315 and 400 nm to form an immobilizing agent covalently bonded to the adhesion promoter layer, wherein the LED UV-A radiation causes a photochemically induced radical chain polymerization of the acrylate monomers and / or acrylate oligomers present in the functional solution and a 3-dimensional polymer structure of the immobilizing agent is formed by covalent crosslinking of the acrylate monomers and / or acrylate oligomers, in and on which reactive groups for immobilizing biomolecules are arranged. Preferably, the adhesion promoter layer in step (a) is applied by means of wet-chemical coating and / or chemical vapor deposition onto at least one surface of the solid substrateapplied. The adhesion promoter layer advantageously comprises predominantly epoxysilane, (3-mercaptopropyl)trimethoxysilane, 3-(trimethoxysilyl)propyl methacrylate, polycyanurates, methacrylate phosphate esters, or isocyanate-based surface additives. In particular, (3-mercaptopropyl)trimethoxysilane, 3-(trimethoxysilyl)propyl methacrylate, and isocyanate-based surface additives can serve as adhesion promoters for the immobilizing agent via chemical interaction with the substrate surface, thus bonding it to the substrate surface, for example via hydrogen bonds or alternatively via covalent addition of the isocyanate group to a Lewis acid hydrogen on the substrate surface. In addition, triazine-containing polycyanurates can form adhesion promoter layers in the form of a thin film, which adsorbs onto the substrate surface through chemical adhesion interactions. In particular, the polycyanurates adsorb through electron interactionsbetween the trioxytriazine rings of the cyanurates and the naturally oxidized Si single crystals of glass substrates (SiO2 on Si (100)). On oxidic aluminum oxide surfaces (Al2O3), the Lewis acidic OH groups serve as electron acceptors, with the lone electron pairs on O and N acting as electron donors. The functional solution is applied to the adhesion promoter layer in step (c) preferably by spin coating, airless spray application, ultrasonic spray application, or inkjet processes. The curing of the functional solution in step (d) is preferably carried out by means of LED UV-A radiation in the wavelength range from 340 to 390 nm or 350 to 390 nm or 360 to 390 nm or 370 to 390 nm or 380 to 390 nm or 385 nm or from 360 to 380 nm or 360 to 370 nm or from 365 nm. By using LED UV-A lamps, in particular in the high-frequency UV-A wavelength range from 340 to 390 nm, many disadvantages of conventional UV-Lamps are avoided. In particular, the formation of harmful ozone is virtually eliminated, so fewer occupational safety measures are necessary and those producing the coating system are exposed to fewer risks. Furthermore, conventional UV lamps are usually mercury-containing lamps, which inherently require greater occupational safety and disposal requirements. LED UV-A lamps often contain specially developed micro-optics for focusing the UV-A radiation, so that more UV-A-mediated energy hits the functional solution, activating many photoinitiators (here, radical generators). In embodiments in which the functional solution is applied using a spin-coating process, curing typically occurs using an LED surface lamp at a wavelength of 365 nm. In this case, the photoinitiator of the functional solution is advantageously a photoinitiator with an absorption maximum of approximately 365 nm.and a comparatively high radical efficiency factor (f). The high radical yield is required for optimal curing of the immobilizing agent. In embodiments in which the functional solution is applied by means of an inkjet process, curing typically takes place using an LED UV lamp integrated into the inkjet printer used at a wavelength of approximately 385 nm. In this case, the photoinitiator of the functional solution is advantageously a photoinitiator with an absorption maximum of approximately 385 nm and, in turn, a comparatively high radical efficiency factor (f). A particular advantage of the inkjet process is that both the process step of applying the functional solution and the curing step take place in one device, namely an inkjet printer. In a further embodiment, the present invention solves the above-mentionedThe aforementioned objects are achieved with a coating system for immobilizing biomolecules on a surface provided with the coating system, comprising at least (a) an adhesion promoter layer and (b) an immobilizing agent covalently bound to the adhesion promoter layer, characterized in that the immobilizing agent has a 3-dimensional polymer structure formed by covalent crosslinking of acrylate monomers and / or acrylate oligomers of a crosslinking degree modulator, wherein the 3-dimensional polymer structure is cured by a radical chain polymerization photochemically induced by UV-A radiation and contains reactive groups for immobilizing biomolecules. In this regard, it should be mentioned that the immobilizing agent of the coating system according to the invention significantly comprises monomers and / or oligomers such as, for example, dipentaerythritol penta- / hexaacrylate, 2-hydroxyethyl 2-methacrylate, glycidyl methacrylate and(Phenylglyoxylic acid methyl ester and diphenyl-(2,4,6-trimethylbenzoyl)phosphine oxide) monomers and / or oligomers. Advantageously, the coating system can be produced particularly cost-efficiently because, in contrast to the use of copolymer-based raw materials, additional, sometimes cost-intensive synthesis, preparation, and analysis steps are eliminated by using monomer- or oligomer-based raw materials. Furthermore, the use of monomer- or oligomer-based raw materials is overall more environmentally compatible and friendly, particularly because monomer- or oligomer-based raw materials can typically be processed directly due to their lower viscosity and chemical structure, whereas copolymer-based raw materials are dissolved in usually toxic organic solvents such as tetrahydrofuran prior to use. The curing of the coating system, and in particular of the immobilizing agentbased on functional acrylate and methacrylate monomers or oligomers is based on the principle of photochemically induced radical chain polymerization. This 05 / 12 / 2024 is usually carried out at room temperature under oxygen conditions or under inert conditions with a residual oxygen content of < 50 ppm by absorbing photons in the wavelength range from 200 nm to 400 nm. The basic prerequisite for radical chain polymerization is the presence of free radicals. While it is technically possible to directly break the π portion of the C=C double bond, this would require a dissociation energy of approximately 6.3 eV at a wavelength of 197 nm. Short-wave radiation of this magnitude would severely limit its application due to ozone formation in the presence of oxygen and a shallow penetration depth of a few micrometers. In order to work in a longer-wavelength spectral range, it is therefore necessaryso-called photoinitiators. It is therefore particularly advantageous that the curing of the coating system according to the invention can be carried out using LED UV lamps due to the use of acrylate monomers or oligomers with free functional acrylate groups. In contrast, the curing of coating systems produced from copolymer-based acrylate raw materials regularly requires the use of mercury-containing UV lamps, as steric hindrances must be overcome. The use of LED UV lamps offers further advantages, such as: ^ Extremely rapid and complete curing of the coating ^ Very little use of volatile solvents required ^ No ozone formation during UV irradiation (high occupational safety, no negative impact on the subsequent coating process) ^ Narrow spectral distribution: Efficiency better than with a mercury-containing UV lamp ^Service life of up to 50,000 hours - UV lamps containing mercury usually have a service life of only 10,000 to 15,000 hours ^ More environmentally friendly: No mercury ^ Low energy consumption ^ Low heat generation: Curing possible with heat-sensitive substrates such as plastic, e.g. PMMA (polymethyl methacrylate). Biomolecules to be immobilized are usually contained in a - often aqueous - sample. 05 / 12 / 2024 The term "biomolecule" in the context of this disclosure includes both high-molecular-weight macromolecules (such as proteins, nucleic acids, polysaccharides or lipids) and low-molecular-weight compounds that are, for example, the building blocks of high-molecular-weight macromolecules (such as amino acids, nucleotides, sugars or fatty acids). Typical biomolecules to be immobilized include nucleic acids, in particular deoxyribonucleic acids (DNA) or ribonucleic acids (RNA); peptides; or proteins, especially enzymes or antibodies.Nucleic acids can in particular be DNA or RNA oligonucleotides. In accordance with the invention, it was initially recognized that the specific binding capacity of a coating system for immobilizing biomolecules can be realized in an astonishingly simple manner by providing the immobilizing agent with a 3-dimensional polymer structure. A 3-dimensional polymer structure is understood to be a structure that results from a 3-dimensional crosslinking of the acrylate monomers and / or oligomers of the crosslinking degree modulator, so that the immobilizing agent – ​​compared to a monolayer of reactive polymers – provides a larger number of reactive groups per unit area and the specific binding capacity of the coating is thus greater. In particular, the immobilizing agent can be designed as a 3-dimensional hydrogel, so that biomolecules contained in a sample can be bound in the 3-dimensional polymer structure by reactionwith the reactive groups are immobilized. The 3-dimensionality of the polymer structure allows the swelling behavior of a coating to be demonstrated by determining the layer thickness using the Quartz Crystal Microbalance and Dissipation (QCM-D) measurement method. According to an advantageous embodiment, the coating system according to the invention for immobilizing biomolecules can be characterized in that: ^ at least some of the acrylate monomers and / or acrylate oligomers of the crosslinking degree modulator are: tetrahydrofurfuryl methacrylate monomers and / or oligomers; and / or 2-phenoxyethyl acrylate monomers and / or oligomers; or ^ at least some of the acrylate monomers and / or acrylate oligomers of the degree of crosslinking modulator contain more than one, preferably more than two or more than three acrylate ester groups per molecule, in particular comprising: trimethylolpropane ethoxy triacrylate monomers and / or 05 / 12 / 2024 oligomers;Dipentaerythritol penta- / hexacrylate monomers and / or oligomers; and / or ^ the immobilizing agent comprises highly branched, functional dendrimers and / or dendrons, preferably highly branched bis(hydroxymethyl)propionic acid, acetylenic, highly branched bis(hydroxymethyl)propionic acid, azide, highly branched bis(hydroxymethyl)propionic acid, hydroxyl and / or highly branched poly(amidoamine) dendrimers and / or dendrons. The immobilizing agent of the coating system according to the invention preferably comprises: ^ an additional raw material, preferably comprising hexanediol diacrylate, poly(ethylene glycol) or polythiol; and / or ^ a hydrophilicity modulator, preferably 2-hydroxyethyl 2-methacrylate; and / or ^ residues of a starting radical of a photoinitiator, preferably of phenylglyoxylic acid methyl ester, of diphenyl-(2,4,6-trimethylbenzoyl)phosphine oxide, of ethyl phenyl(2,4,6-trimethylbenzoyl)phosphinate or of phenyl bis(2,4,6-trimethylbenzoyl)phosphine oxide; and / or ^ aAn immobilization modulator that provides the reactive groups for immobilizing biomolecules. The immobilization modulator preferably comprises: ester groups, preferably N-hydroxysuccinimide ester groups, glycidyl ester groups, or isocyanate ester groups; ether groups, preferably glycidyl ether groups; epoxy groups; aldehyde groups; free carboxyl and / or free amino groups, and combinations thereof. Furthermore, the 3-dimensional polymer structure of the immobilizing agent is preferably obtained by curing a functional solution comprising: ^ between 12 and 32 weight percent of the crosslinking degree modulator; ^ between 11 and 16 weight percent of the hydrophilicity modulator; ^ between 7 and 18 weight percent of the additional raw material; ^ between 20 and 50 weight percent of the immobilization modulator; and ^ between 3.5 and 6.5 weight percent of the photoinitiator. Particularly advantageously, the UV-A radiation is an LED-UV-ARadiation in the wavelength range of 315 to 400 nm, preferably in the wavelength range of 340 to 390 nm or 350 to 390 nm or 360 to 390 nm or 370 to 390 nm or 380 to 390 nm or 385 nm or 360 to 380 nm or 360 to 370 nm or 365 nm. By using LED UV-A lamps, particularly in the high-frequency UV-A wavelength range of 340 to 390 nm, many of the disadvantages of conventional UV lamps are avoided. Since LED UV lamps emit UV-A energy, contain no mercury, produce no heat and produce practically no ozone, fewer occupational safety measures are necessary overall and people producing the coating system are exposed to significantly lower risks. Preferably, the adhesion promoter layer comprises predominantly derivatives of epoxysilane, (3-mercaptopropyl)trimethoxysilane, 3-(trimethoxysilyl)propyl methacrylate, polycyanurates, methacrylate phosphate esters or isocyanate-based surface additives which are covalently bonded withAcrylate monomers and / or acrylate oligomers of the immobilizing agent. Particularly advantageously, the binding reaction covalently with the acrylate monomers and / or oligomers and, for example, an amino or thiol group of a biomolecule does not produce a condensation product that could remain in the coating system. Advantageously, the adhesion promoter layer can be applied to a surface by chemical vapor deposition. Alternatively, the adhesion promoter layer can be applied to a suitably prepared substrate surface (for example, to a cleaned and activated glass substrate surface) by means of a wet-chemical coating process, in particular a dipping, pouring, spraying, or pressure coating process. Both application by chemical vapor deposition and by a wet-chemical coating process enables the rapid and cost-effective production of athe adhesion promoter layer. Suitable and already known methods for applying the immobilizing agent to the adhesion promoter layer are spin coating, slot-die coating, airless spray application, ultrasonic spray application, or inkjet methods. In particular, the use of an immobilizing agent that is curable by means of UV-A radiation photochemically induced radical polymerization enables the particularly advantageous application of the immobilizing agent by the inkjet method. The inkjet method enables the production of microarrays and biochips provided with the coating system according to the invention in a particularly efficient and simple manner. 05 / 12 / 2024 The inventive combination of an adhesion promoter layer with the immobilizing agent provides a coating system, wherein the contact angle of water on the immobilizing agent is optionally greater than 60°, preferably betweenbetween 60° and 70°. Due to the preferably adjusted contact angle, relatively large volumes of aqueous sample solutions can be applied in spots with a small diameter, for example a diameter of between 140 and 170 µm, to the surface provided with the coating system, which in turn ensures a high concentration of biomolecules to be analyzed per spot. The combination of such a large number of biomolecules per spot and the advantageously high binding capacity of the immobilizing agent leads to a high signal intensity of the biomolecules to be analyzed in the subsequent analysis methods, which significantly facilitates their evaluation. Preferably, the coating system is solvent-resistant to organic solvents, in particular to acetone, toluene, or methanol, so that after treatment with organic solvents, in particular after treatment with toluene or methanol for 30minutes at 60°C, shows no detectable damage. This robustness of the coating system enables its use in a wide variety of analytical methods. In a further embodiment, the present invention achieves the above-mentioned objects by means of a solid-state substrate for the immobilization of biomolecules, which is produced using the aforementioned inventive method and comprises at least one surface provided with the aforementioned inventive coating system, wherein the substrate is preferably a glass substrate or plastic substrate or a silicon substrate or an oxidic substrate and is suitable for the production of a microarray or a biochip. The solid-state substrate is preferably a microarray or biochip for the immobilization of biomolecules. As already mentioned, the biomolecules to be immobilized for analysis are usually contained in a - often aqueous - sample.Typical biomolecules to be analyzed include nucleic acids, especially deoxyribonucleic acids (DNA) or ribonucleic acids (RNA); peptides; or proteins, especially enzymes. Nucleic acids to be analyzed can, in particular, be DNA or RNA oligonucleotides. 05 / 12 / 2024 Excitable dyes, which emit detectable radiation upon excitation, are typically used to label and detect immobilized biomolecules. Suitable fluorophores include, for example: Name Absorbing Emitting Wavelength Visible Color Wavelength Hydroxycoumarin 325 386 Blue methoxycoumarin 360 410 Blue Alexa fluor 345 442 Blue aminocoumarin 350 445 Blue Cy2 490 510 Dark green FAM 495 516 Dark green Alexa fluor 488 494 517 Light green Fluorescein FITC 495 518 Light green Alexa fluor 430 430 545 Light green Alexa fluor 532 530 555 Light green HEX 535 556 Light green Cy3 550 575 Yellow TRITC 547 572 Yellow Alexa fluor 546 556 573 Yellow Alexa fluor 555 556573 Yellow R-phycoerythrin (PE) 480;565 578 Yellow Rhodamine Red-X 560 580 Orange Tamara 565 580 Red Cy3.5581 581 596 Red Rox 575 602 Red Alexa fluor 568 578 603 Red Red 613 480;565 613 Red Texas Red 615 615 Red Alexa fluor 594 590 617 Red Alexa fluor 633 621 639 Red Allophycocyanin 650 660 Red Alexa fluor 633 650 668 Red Cy5 650 670 Red 05 / 12 / 2024 Alexa fluor 660 663 690 Red Name Absorbing Emitting Wave Visible Color Wavelength Cy5.5 675 694 Red TruRed 490;675 695 Red Alexa fluor 680 679 702 Red Cy7 743 770 Red Particularly suitable for labeling DNA or RNA molecules are cyanine fluorophores, including the known fluorophores Cy2, Cy3, Cy3B, Cy3.5, Cy5, Cy5.5 or Cy7. Preferably, a microarray or biochip comprising a surface provided with the aforementioned coating system according to the invention has only a low background intensity. This low background intensity ensures thatEmitted signals in the wavelength range of approximately 500 nm to 575 nm can be detected essentially without interfering background signals from the substrate itself. Advantageously, the sensitivity of a microarray or biochip produced on the basis of the solid-state substrate according to the invention is thus high. Those skilled in the art know how the background intensity is determined in relation to the signal intensity to be determined in a fluorescence-based measurement system. For example, a microplate reader from TECAN can be used for this purpose, in which both the background intensity and the signal intensity to be determined of a microarray or biochip produced on the basis of the solid-state substrate according to the invention are determined via an optical system. In this system, the substrate surface is first irradiated by a light source with a wavelength of 532 nm in excitation mode. This leads to fluorescence emitted by the glass surface.The relevant emission wavelength is selected using an optical filter, for example, with a wavelength of 575 nm. The intensity of the emitted light can then be determined using a detector. In such fluorescence measurements, a signal amplification "gain" can be set as required, which can, of course, strongly influence fluorescence measurements. In this respect, the person skilled in the art knows that the definition of this parameter is particularly important in every measurement system in order to be able to determine a low background intensity compared to the signal intensity to be determined. 05 / 12 / 2024 A solid-state substrate that can be provided with the coating system according to the invention for use as a microarray or biochip emits only a very low fluorescence signal before the coating system is applied. If, after the application of the adhesion promoter layer, upon excitation with a wavelength of 532 nm, aIntrinsic fluorescence with a signal strength of approximately 40±10 relative fluorescence units (rfu) is determined, the substrate is also suitable for the application of the immobilization agent, since the background signal strength of the ready-to-use microarray or biochip, which results from the intrinsic fluorescence signal of the coated substrate and the background signal generated by non-specific binding or immobilization of dye, is then usually still significantly less than 150 rfu after excitation with a wavelength of 532 nm. Example 1 shows that (a) the intrinsic fluorescence of a suitable glass substrate is 22 ± 0.5 rfu, (b) the intrinsic fluorescence of the glass substrate with the adhesion promoter layer applied by chemical vapor deposition is 26.2 ± 1.0 rfu, and (c) the intrinsic fluorescence of the glass substrate with the coating system according to the invention is 68.4 ± 1.4 rfu. The intrinsic fluorescence of aIn direct comparison, the value of known microarrays equipped with a conventional coating system is 172.8 ± 20.1 rfu, which is more than 2.5 times higher. With regard to a plastic substrate to be used in the aforementioned solid-state substrates, microarrays, or biochips, this can be a polymethyl methacrylate substrate (PMMA substrate). With regard to a glass substrate to be used in the aforementioned solid-state substrates, microarrays, or biochips, it is conceivable that this is selected from: soda-lime glass, borosilicate glass, quartz glass, and / or alkali-free alumino-borosilicate glass. Preferably, the glass of a glass substrate to be used in the solid-state substrate according to the invention can have the following composition components corresponding to a lithium aluminum silicate glass (in weight %): SiO255-69 Al2O318-25 Li2O 3-5 Na2O+K2O 0-30 05 / 12 / 2024 MgO+CaO+SrO+BaO 0-5 ZnO 0-4 TiO20-5 ZrO20-5 TiO2+ZrO2+SnO22-6 P2O50-8 F 0-1 B2O30-2Again, the glass of a glass substrate to be used in the solid-state substrate according to the invention can preferably have the following components (in % by weight): SiO257-66 Al2O318-23 Li2O 3-5 Na2O+K2O 3-25 MgO+CaO+SrO+BaO 1-4 ZnO 0-4 TiO2O-4 ZrO2O-5 TiO2+ZrO2+SnO22-6 P2O5O-7 F 0-1 B2O3O-2

[0002] 05 / 12 / 2024 Again preferably, the glass of a glass substrate to be used in the solid-state substrate according to the invention can have the following components (in weight %): SiO257-63 Al2O318-22 Li2O 3.5-5 Na2O+K2O 5-20 MgO+CaO+SrO+BaO 0-5 ZnO 0-3 TiO2O-3 ZrO2O-5 TiO2+ZrO2+SnO22-5 P2O50-5 F 0-1 B2O30-2 Again preferably, the glass of a glass substrate to be used in the solid-state substrate according to the invention can have the following components corresponding to a soda-lime silicate glass (in weight %): SiO240-81 Al2O30-6 B2O30-5 Li2O+Na2O+K2O 5-30 MgO+CaO+SrO+BaO+ZnO 5-30 TiO2+ZrO2O-7 P2O5O-2 Again, the glass of a glass substrate to be used in the solid substrate according to the invention can preferably have the following components (in weight %): 05 / 12 / 2024 SiO25O-81 Al2O3O-5 B2O3O-5 Li2O+Na2O+K2O 5-28 MgO+CaO+SrO+BaO+ZnO 5-25 TiO2+ZrO2O-6 P2O5O-2 Again, the glass of a glass substrate to be used in the solid substrate according to the invention can preferably have the following componentsComponents (in weight %): SiO250-76 Al2O30-5 B2O30-5 Li2O+Na2O+K2O 5-25 MgO+CaO+SrO+BaO+ZnO 5-20 TiO2+ZrO20-5 P2O50-2 Again preferably, the glass of a glass substrate to be used in the solid substrate according to the invention can have the following components corresponding to a borosilicate glass (in weight %): SiO260-85 Al2O30-10 B2O35-20 Li2O+Na2O+K2O 2-16 MgO+CaO+SrO+BaO+ZnO 0-15 TiO2+ZrO20-5 P2O50-2 Again preferably, the glass of a glass substrate to be used in the solid substrate according to the invention can have the following components (in Weight-%): 05 / 12 / 2024 SiO263-84 Al2O30-8 B2O35-18 Li2O+Na2O+K2O 3-14 MgO+CaO+SrO+BaO+ZnO 0-12 TiO2+ZrO20-4 P2O50-2 Again preferably, the glass of a glass substrate to be used in the solid-state substrate according to the invention can have the following components (in weight-%): SiO263-83 Al2O30-7 B2O35-18 Li2O+Na2O+K2O 4-14 MgO+CaO+SrO+BaO+ZnO 0-10 TiO2+ZrO20-3 P2O50-2 Again preferably, the glass of aThe glass of a glass substrate to be used in the solid-state substrate according to the invention can have the following components (in weight %): SiO260-70 Al2O31-10 B2O31-10 K2O 1-10 Na2O 1-10 ZnO 1-10 TiO21-10 The glass of a glass substrate to be used in the solid-state substrate according to the invention can preferably have the following components corresponding to an alkali aluminum silicate glass (in weight %): 05 / 12 / 2024 SiO240-75 Al2O310-30 B2O30-20 Li2O+Na2O+K2O 4-30 MgO+CaO+SrO+BaO+ZnO 0-15 TiO2+ZrO20-15 P2O50-10 The glass of a glass substrate to be used in the solid-state substrate according to the invention can preferably to be used glass substrate have the following components corresponding to a low-alkali aluminum silicate glass (in weight %): SiO250-70 Al2O310-27 B2O30-18 Li2O+Na2O+K2O 5-28 MgO+CaO+SrO+BaO+ZnO 0-13 TiO2+ZrO20-13 P2O50-9 Again preferably, the glass of a glass substrate to be used in the solid-state substrate according to the invention can have the following components (in weight %): SiO255-68Al2O310-27 B2O30-15 Li2O+Na2O+K2O 4-27 MgO+CaO+SrO+BaO+ZnO 0-12 TiO2+ZrO20-10 P2O50-8 Again, the glass of a glass substrate to be used in the solid-state substrate according to the invention can preferably have the following components (in weight %): 05 / 12 / 2024 SiO250-75 Al2O37-25 B2O30-20 Li2O+Na2O+K2O 0-4 MgO+CaO+SrO+BaO+ZnO 5-25 TiO2+ZrO20-10 P2O50-5 Again, the glass of a glass substrate to be used in the solid-state substrate according to the invention can preferably have the following components (in weight %): SiO252-73 Al2O37-23 B2O30-18 Li2O+Na2O+K2O 0-4 MgO+CaO+SrO+BaO+ZnO 5-23 TiO2+ZrO20-10 P2O50-5 Again, the glass of a glass substrate to be used in the solid substrate according to the invention can preferably have the following components (in weight %): SiO253-71 Al2O37-22 B2O30-18 Li2O+Na2O+K2O 0-4 MgO+CaO+SrO+BaO+ZnO 5-22 TiO2+ZrO20-8 P2O50-5 It is understood that the respective glass components of the listed glass compositions total 100% by weight.must. Nevertheless, the glasses to be used in the invention, in particular the glasses described above, can in turn be modified. For example, the color of the respective glass can be changed by adding color oxides. 05 / 12 / 2024 In advantageous embodiments, the glass substrates according to the invention are produced using particularly pure raw materials in order to minimize fluorescence under illumination with UV radiation and / or radiation in visible light. In particular, the use of raw materials with a very low iron content has proven advantageous for this purpose. The glasses produced in this way therefore advantageously contain particularly few impurities, in particular little iron. In a further embodiment, the present invention achieves the aforementioned objects with a microarray or biochip for the immobilization of biomolecules, preferably for the analysis of biomolecules contained in a sample.comprising the coating system according to the invention and / or the solid-state substrate according to the invention, wherein the biomolecules are preferably: nucleic acids, in particular deoxyribonucleic acids (DNA) or ribonucleic acids (RNA); peptides; or proteins, in particular enzymes. In a further embodiment, the present invention achieves the above-mentioned objects with a functional solution for use in a method according to the invention and / or for use in the production of an immobilizing agent of the coating system according to the invention, at least comprising: ^ a crosslinking degree modulator comprising: acrylate monomers and / or acrylate oligomers with one acrylate ester group per molecule, preferably tetrahydrofurfuryl methacrylate monomers and / or oligomers; and / or 2-phenoxyethyl acrylate monomers and / or oligomers; and / or acrylate monomers and / or acrylate oligomers with more than one, preferably more than two or more than threeContaining acrylate ester groups per molecule, in particular comprising: trimethylolpropane ethoxy triacrylate monomers and / or oligomers; dipentaerythritol penta / hexacrylate monomers and / or oligomers; and / or highly branched functional dendrimers and / or dendrons, preferably highly branched bis(hydroxymethyl)propionic acid, acetylenic, highly branched bis(hydroxymethyl)propionic acid, azide and / or highly branched bis(hydroxymethyl)propionic acid, hydroxyl and / or highly branched poly(amidoamine) dendrimers and / or dendrons; ^ an immobilization modulator which provides reactive groups for the immobilization of biomolecules, the immobilization modulator preferably comprising: ester groups, preferably N-hydroxysuccinimide ester groups, glycidyl ester groups or isocyanate ester groups; ether groups, preferably glycidyl ether groups; epoxy groups; aldehyde groups; free carboxyl and / or free amino groups, as well as combinations thereof; and ^ aPhotoinitiator with an absorption maximum in the wavelength range between 315 and 400 nm, preferably a photoinitiator comprising: phenylglyoxylic acid methyl ester and diphenyl-(2,4,6-trimethylbenzoyl)phosphine oxide; ethyl phenyl(2,4,6-trimethylbenzoyl)phosphinate; phenyl bis(2,4,6-trimethylbenzoyl)phosphine oxide; or 2-hydroxy-2-methylpropiophenone. Highly branched, functional dendrimers and / or dendrons preferably comprise highly branched bis(hydroxymethyl)propionic acid acetylene, highly branched bis(hydroxymethyl)propionic acid azide and / or highly branched bis(hydroxymethyl)propionic acid hydroxyl and / or highly branched poly(amidoamine) dendrimers and / or dendrons, such as ^ Bis(hydroxymethyl)propionic acid acetylene dendrimers (Bis-MPA acetylene dendrimers; Polymer Factory; Product Code: PFD-G1-TMP-ACETYLENE) are monodisperse, multifunctional frameworks with a trimethylolpropane (TMP) core and a precise number of surface groups. TheseAlkyne-functional dendrimers can be used for various click chemistry reactions such as CuAAC and thiol-yne coupling; ^ Bis(hydroxymethyl)propionic acid azide dendrimers (Bis-MPA azide dendrimers; Polymer Factory; Product Code: PFD-G1-TMP-AZIDE) are monodisperse, multifunctional scaffolds with a trimethylolpropane (TMP) core and a precise number of azide surface groups. The azide-functional dendrimers can also be used for click chemistry reactions such as CuAAC or SPAAC. ^ Bis(hydroxymethyl)propionic acid hydroxyl dendrons (Bis-MPA hydroxyl dendrons; Polymer Factory; Product Code: PFd-G2-Acetylene-OH) are monodisperse, multifunctional scaffolds with a central acetylene group and a precise number of hydroxyl surface groups. The central acetylene group can be used for click chemistry conjugations such as CuAAC and thiol-yne coupling to produce highly functional materials; ^ a highly branched poly(amidoamine) which serves asIt is considered analogous to PAMAM dendrimers. It has primary amine end groups and carboxylic acids, which enables a variety of reactions for easy modification or network formation. 05 / 12 / 2024 Bis(hydroxymethyl)propionic acid polyester and poly(amidoamine) scaffolds are biodegradable and exhibit low cytotoxicity. The aforementioned functional solution preferably further comprises: ^ an additional raw material, preferably comprising hexanediol acrylate, poly(ethylene glycol), or polythiol; and / or ^ a hydrophilicity modulator, preferably 2-hydroxyethyl 2-methacrylate. The crosslinking degree modulators specified here allow the adjustment of the degree of crosslinking of the acrylate monomers and / or oligomers within the functional solution achieved by the radical chain polymerization photochemically induced by UV-A radiation. In combination with the immobilization modulators, which provide the reactive groups for interaction with theThe functionality, structure, and concentration of the selected crosslinking and immobilization modulators can be used to adjust the solvent and temperature resistance, scratch resistance, polymer shrinkage, and the direct adjustment of the background intensity, as well as the binding capacity of the 3-dimensional immobilizing agent. In particular, when crosslinking modulators with more than one, preferably more than two or more than three acrylate ester groups per molecule are used, the 3-dimensional polymer structure of the immobilizing agent can be advantageously adjusted. It is important to ensure that a balance is found between the resistance of the immobilizing agent and the binding capacity of the immobilizing agent for biomolecules. For example, the binding capacity of the immobilizing agent decreases when usingThe degree of crosslinking modulators decreases with increasing branching, while the degree of crosslinking increases. As the degree of crosslinking increases, the immobilization agent becomes more compact and the accessibility of biomolecules to the reactive groups in the coating decreases. In order to induce photochemical radical chain polymerization of the acrylate monomers or oligomers using UV-A radiation, the functional solution also includes a photoinitiator. The photoinitiator is particularly characterized by its high yield in the generation of free radicals upon irradiation with UV-A radiation in the wavelength range between 315 and 400 nm. Photoinitiators are known and 05 / 12 / 2024 a preferred, commercially available photoinitiator comprises phenylglyoxylic acid methyl ester and diphenyl-(2,4,6-trimethylbenzoyl)-phosphine oxide (5.8 wt%) and has an absorption maximum at a wavelength of approximately 365 nm. An alternative but equally suitablePhotoinitiator comprises ethyl phenyl(2,4,6-trimethylbenzoyl)phosphinate and has an absorption maximum at a wavelength of approximately 385 nm. Another suitable photoinitiator comprises phenylbis(2,4,6-trimethylbenzoyl)phosphine oxide and has an absorption maximum at a wavelength of approximately 370 nm. In particular embodiments in which the functional solution comprises an additional raw material, the additional raw material preferably serves to determine the flexibility of the immobilizing agent after curing. In particular, the additional raw material leads to advantageous shrinkage of the immobilizing agent during curing. The additional raw material, through its swelling behavior, supports the 3-dimensional polymer structure of the immobilizing agent and can simultaneously prevent the nonspecific binding of other components of the sample. In addition, the reduction of nonspecific binding is achieved by increasing the polar portion.of the free surface energy is achieved. By increasing the polarity of the coating, hydrophobic interactions between the biomolecules and the coating are prevented. To adjust the contact angle of water on the immobilizing agent so that it is optionally greater than 60°, preferably between 60° and 70°, the functional solution preferably additionally comprises a hydrophilicity modulator. The hydrophilicity modulator further improves the adhesion of the coating system to the surface and reduces the non-specific binding of biomolecules. In some embodiments, the substrate adhesion of the coating system is achieved due to the formation of hydrogen bonds between free SiOHx groups and hydroxyl groups of the hydrophilicity modulator 2-hydroxyethyl-2-methacrylate. The functional solution preferably comprises: ^ between 12 and 32 weight percent of the crosslinking degree modulator; ^ between 11 and 16Weight percent of the hydrophilicity modulator; ^ between 7 and 18 weight percent of the additional raw material; ^ between 20 and 50 weight percent of the immobilization modulator; and ^ between 3.5 and 6.5 weight percent of the photoinitiator. 05 / 12 / 2024 As already described, the aforementioned solid-state substrate is preferably a microarray or biochip for immobilizing biomolecules. As also already described, the biomolecules to be immobilized on the solid-state substrates according to the invention are preferably nucleic acids, in particular deoxyribonucleic acids (DNA) or ribonucleic acids (RNA); peptides; or proteins, in particular enzymes. The process according to the invention enables the production of large quantities of the solid-state substrates, microarrays and biochips according to the invention with particularly high resistance to aqueous solutions and solvents such as methanol, toluene and / or acetone, even at increasedTemperature. In a further embodiment, the present invention achieves the above-mentioned objects by using a coating system according to the invention for immobilizing biomolecules on a surface of a microarray or a biochip provided with the coating system, wherein the biomolecules are preferably: nucleic acids, in particular deoxyribonucleic acids or ribonucleic acids; peptides; or proteins, in particular enzymes. There are now various possibilities for advantageously embodying and developing the teaching of the present invention. Reference is made, on the one hand, to the claims subordinate to claim 1 and, on the other hand, to the following explanation of preferred embodiments of the invention with reference to the drawings. In conjunction with the explanation of the preferred embodiments of the invention with reference to the drawings, generally preferred embodiments and developments are also described.Further developments of the teaching are explained. In the drawings, Fig. 1 schematically illustrates the steps of the inventive production method of a coating system on the surface of a solid substrate. Fig. 2 shows a bar chart in which - according to Example 2 - the polar component of the free surface energy and the fluorescence intensity of an immobilized fluorescently labeled protein is shown as a function of the concentration of 2-hydroxyethyl-2-methacrylate (HEMA) in the immobilizing agent. Fig.3 shows - according to Example 3 - the transmission spectrum of a cleaned solid substrate (here a glass slide; Sample 1), a solid substrate with 05 / 12 / 2024 the adhesion promoter layer of the coating system according to the invention (here a corresponding glass slide with a 3-(trimethoxysilyl)propyl methacrylate-based adhesion promoter layer; Sample 2), a solid substrate with the coating system according to the invention (Sample 3) andof a corresponding solid-state substrate with a known copolymer-based coating system (sample 4). Fig. 4 shows - according to Example 4 - representative light micrographs (20x magnification) of the coating areas of a glass substrate surface provided with the coating system according to the invention after aging in solvent: (a) after aging for 48 h at room temperature RT in toluene; (b) after aging for 48 h at room temperature RT in methanol; and (c) after aging for 48 h at room temperature RT in acetone. Fig. 5 shows - according to Example 5 - the results of cross-cut tests concerning a known coating system applied to a glass substrate (comparative sample) and two samples of the coating system according to the invention applied to corresponding glass substrates; OK = OK; NOK = NOT OK. Fig. 6 shows - according to Example 6 - the results of the contact angle measurement of water onAdhesion promoter layers based on 3-methacryloxypropyltrimethoxysilane and as a function of silane concentration. Figure 7 shows - according to Example 6 - the results of the background intensity measurements of adhesion promoter layers based on 3-methacryloxypropyltrimethoxysilane and as a function of silane concentration and temperature. Figure 8 shows - according to Example 7 - a schematic representation of the DNA molecules bound in the 3-dimensional polymer structure of the immobilizing agent upon hybridization of a DNA sample to be analyzed. Figure 9 shows - according to Example 7 - the results of a comparative study of the oligonucleotide binding capacity of a coating system according to the invention (with a 3D polymer structure) and a 2D epoxy coating as a function of concentration. Figure 10 shows - according to Example 8 - results of the measurement of the nonspecific binding of proteins on a coating system according to the invention (with a 3D polymer structure) in comparison.to a 2D epoxy coating. 05 / 12 / 2024 Fig. 11 shows - according to Example 8 - results of the measurement of the non-specific binding of proteins on a coating system according to the invention (with a 3D polymer structure) in comparison to a 2D epoxy coating. Fig. 12 shows - according to Example 9 - the relationship between the increased contact angle of water on a coating system according to the invention and the spot diameter of an applied sample. Fig. 13 shows - according to Example 9 - the increased binding capacity of the coating system according to the invention per spot. Regarding further advantageous embodiments of the coating system according to the invention for the immobilization of biomolecules, the solid-state substrate according to the invention for the immobilization of biomolecules, the microarray or biochip according to the invention for the immobilization of biomolecules, the functional solution according to the invention for producing an immobilization agent, and the method according to the inventionTo avoid repetition, reference is made to the general part of the description as well as to the appended examples and claims. Figure 1 schematically illustrates the steps of the inventive production method of a coating system (1) on a surface (2) of a solid substrate (3). First, an adhesion promoter layer (4) comprising methacrylateorganosilane or thiolorganosilane monomers is applied to a surface (2) of a glass substrate (3), preferably by chemical vapor deposition (CVD). Catalyzed hydrolysis and condensation of the silyl ether compound of the methacrylate or thiolsilane adsorbed on the surface (2) leaves free (i.e., reactive) acrylate groups (5) or free (i.e., reactive) thiol groups (6) when using methacrylateorganosilane monomers or thiolorganosilane monomers (Figure 1(a)).Trimethoxy or triethoxysilyl groups bind the methacrylate organosilane or thiol organosilane monomers to the surface of the glass substrate. These reactive groups (5, 6) of the adhesion promoter layer can form covalent bonds with components of the immobilizing agent (7) to be applied, thus mediating the optimal adhesion of the immobilizing agent (7) to the adhesion promoter layer (4) on the one hand and indirectly to the surface (2) of the glass substrate (3) on the other. In the next step, the prepared functional solution is applied to the adhesion promoter layer and cured by photochemically induced radical chain polymerization using LED UV-A radiation in the wavelength range between 315 and 400 nm. In particular, an intermolecular cross-linking takes place between the reactive groups (5,6) of the adhesion promoter layer (4) with components of the functional solution so that the 3-dimensional polymer structure of theImmobilizing agent forms a layer in and on which reactive epoxy groups (8) are arranged. The reactive epoxy groups (9) can react with amino groups of the biomolecules to be immobilized through epoxy opening, thereby immobilizing the biomolecules. Examples The composition of immobilizing agents according to the invention can be found in the following table: Recipe 2.5 Components Percentage (wt%) Dipentaerythritol penta- / hexaacrylate 21.2 2-Hydroxyethyl 2-methacrylate 13.5 Poly(ethylene glycol) 15.4 Glycidyl methacrylate 44.2 Methyl phenylglyoxylate and diphenyl-(2,4,6-trimethylbenzoyl)phosphine oxide (5.8 wt%) or 5.8 Ethyl phenyl(2,4,6-trimethylbenzoyl)phosphinate SUM: 100 Recipe 2.9 Components Percentage (wt%) (Hexanediol diacrylate) HDDA (Flexibility) 10.4 Trimethylolpropane ethoxy triacrylate 29.8 Glycidyl methacrylate 41.8 2-Hydroxyethyl-2-methacrylate 14.0 2,4,6-Trimethylbenzoyldiphenylphosphine 4.0 oxide TOTAL: 10005 / 12 / 2024 Recipe 3.9 Ingredients Percentage (wt%) Tetrahydrofurfuryl acrylate 11 - 12 Trimethylolpropane ethoxy triacrylate 41 - 54 Glycidyl methacrylate 14 - 27 2-Hydroxyethyl-2-methacrylate 14 Ethyl phenyl(2,4,6-trimethylbenzoyl)phosphinate 6 - 8 SUM: 100 Example 1 Comparative autofluorescence determination: (a) of a suitable glass substrate (BOROFLOAT 33, thickness 1.0 ± 0.05 mm; SCHOTT AG, Mainz, Germany, (b) of the glass substrate provided with a methacryloxypropyltrimethoxysilane-based adhesion promoter layer by chemical vapor deposition (CVD), (c) of the glass substrate provided with a coating system according to the invention, wherein the coating system by means of CVD with a methacryloxypropyltrimethoxysilane-based adhesion promoter layer and an immobilizing agent according to formulation 2.5, and (d) a commercially available microarray comprising a conventional coating system (TRIDIA HD, Surmodics IVD, Inc, Eden Prairie, USA).Determination procedure carried out as follows: G erät:Tecan Scanner 400 TECAN Germany GmbH Excitation wavelength 532 nm 575 nm Emission wavelength Instrument settings (Cy3 filter): Signal amplification_PMT 200 (photomultiplier tube) Gain Scan resolution 10 µm Measurement results 05 / 12 / 2024 Fluorescence Standard Variation Material intensity deviation coefficient [rfu] (a) BOROFLOAT 33 22 0.5 0.02 (b) BOROFLOAT 33 with adhesion promoter 26.2 1.0 0.04 layer (c) BOROFLOAT 33 with coating system 68.4 1.4 0.02 (d) TRIDIA HD 172.8 20.1 0.12 Example 2 In general, the optimal functionality of a microarray and its evaluation depends The binding behavior of the coating system on the solid substrate depends significantly on its own fluorescence and the specific binding behavior of the coating system of the microarray.To reduce nonspecific binding and thus an increased background signal after incubation with a sample, the immobilization agent was modified by incorporating the raw material 2-hydroxyethyl-2-methacrylate (HEMA) to gradually increase the polar portion of the free surface energy and investigated using NSB tests (NSB = non-specific binding). The specific and nonspecific binding behavior of a coating system is tested using an NSB test, in which a glass substrate coated with the coating system is incubated with a fluorescently labeled protein mix (goat serum - DY-547P1). By increasing the polar portion of the free surface energy through the incorporation of HEMA, lower nonspecific binding on the coating system can be achieved. The intensities shown in Figure 2 were normalized against the intensities of the cleaned glass substrate without the coating system (negative control).Material: - goat serum-DY-547P1 - 1xPBS / 0.1% Tween 20 - Deionized water 05 / 12 / 2024 Results: As can be seen from an experiment, a coating system without a hydrophilicity modulator (here without HEMA; negative control) exhibits very strong nonspecific binding behavior. Advantageously, the signal intensity of the immobilized fluorescently labeled protein decreases continuously with increasing HEMA concentration in the immobilizing agent. The decrease in the signal intensity of the immobilized fluorescently labeled protein is presumably due to the reduction in hydrophobic protein-solid interactions. The coating surface can enter into different interactions with the adsorbed proteins on both a microscopic and macroscopic scale and can also influence their conformation.First, precisely those factors that strongly influence the properties of the solid surface should be considered. An important driving force for protein adsorption is the hydrophobic interaction with the solid surface. In general, hydrophobic properties make the greatest contribution to protein adsorption compared to electrostatic properties and depend on surface polarity. In this context, studies on the resistance to protein adsorption in oligoether-based self-assembled monolayers (SAMs) using factors such as internal hydrophilicity, terminal hydrophilicity, and lateral density of the SAM layer have shown that increasing hydrophobicity of the oligo(ethylene glycol) layer due to the introduced hydrophobic end groups increases the adsorption of the protein fibrinogen.In summary, immobilization depends on the following parameters: - the physical nature of the support (e.g. porosity, shape, etc.) - the chemical nature of the support (chemical composition, in particular cross-linking, branching, number of functional groups, reactivity of the functional groups, etc.) - the nature of the linkage or binding chemistry - the conformation of the protein during and after immobilization; the type and length of the spacer - the properties of the medium during attachment of the selected protein - the individual properties of the selected protein; - the number of bonds between the protein and the support - the distribution of the protein on or in the support.05 / 12 / 2024 Example 3 To enable the measurement of signal intensities of immobilized fluorescently labeled biomolecules through a glass substrate coated with the coating system, the "LED-UV coating" coating system must exhibit high transparency. Since UV coating systems are cured using short-wave radiation, it can be assumed that they are at least partially transparent to ultraviolet radiation, otherwise the radiation cannot propagate through the bulk layer. In this context, the question of the transparency and UV activity of the coating should be clarified by measuring the absorbance and transmittance at the respective wavelength ranges in the UV / Vis spectrum. Materials: A SPECORF200 spectrometer from Analytik Jena was used, and the measured transmission spectra are shown in Figure 2.Results: As can be seen from the results shown in Figure 3, the application of the coating system according to the invention to a glass substrate does not lead to an unfavorable restriction of the transmission (see transmission sample 3 in Figure 2). In particular, the coating system according to the invention (sample 3 in Figure 2) has comparably high transmission values ​​as a coating system already available on the market (sample 4 in Figure 2). Example 4 Since organic solvents such as toluene, methanol, or acetone are used as fixatives in cytodiagnostics, a favorable coating system must have good solvent resistance. To test the solvent resistance of the coating system according to the invention, the following steps were carried out: - Step 1: Before storage - scanning using a Tecan scanner (control) - Step 2: KW measurement using a contact angle measuring device from Krüss - Step 3:3: Exposure of substrates in the following organic solvents such as toluene, methanol, and acetone at room temperature (RT) for 48 hours. - Step 4: After exposure, dry in a nitrogen stream on 05 / 12 / 2024. - Step 5: After the drying process, dry in a substrate holder or rack in an oven at a temperature of 30°C for 30 minutes. - Step 6: Cool to RT for 20 minutes. Step 7: Measure background fluorescence using a Tecan scanner (result). - Step 8: Light microscopic (LM) assessment of durability: Take LM images of treated and untreated areas at 2.5x and 20x magnification. The coating should be intentionally damaged with the glass pen so that the coating side can also be visually identified in the image. - Step 9: Contact angle measurements (CA measurements) on the treated and untreated areas. - Step 10: Documentation of the test procedure, includingEvaluation of results: As shown in Figures 4a) to 4c), the coating system according to the invention exhibits very good solvent resistance to toluene, methanol, and acetone at room temperature. Example 5 In order to test the adhesion strength of the coating system to the solid surface, the formulated coating was subjected to a cross-cut test in accordance with DIN EN ISO 2409. The requirement for the coating with regard to adhesion strength was to achieve a cross-cut characteristic value of 0 - 1. The cross-cut test according to DIN EN ISO 2409 is one of the most widely used tests of coating surfaces for determining the adhesion strength of a coating to the substrate. It is easy to carry out and does not require a great deal of equipment or time. The coating surface is scratched down to the substrate using a craft knife. The sample is then peeled off by approx.The sample is rotated by 90° and the process is repeated. This creates a scratched grid pattern on the sample. For hard substrates, an adhesive strip is then placed parallel to a cut edge over the grid pattern and pressed down. After a maximum of 5 minutes, the adhesive tape is peeled off evenly within 0.5 seconds to 1.0 seconds at an angle of 60°. The resulting cross-cut is then examined with a magnifying glass and documented by a light microscope image. There are six cross-cut parameters. 05 / 12 / 2024 Cross-cut parameter 0 (GT0) means that the pattern has remained completely unchanged and no detachment or fraying of the cut edges can be observed at any point.With a cross-cut characteristic value of 5, more than 65% of the cross-cut area has been detached. In principle, poor adhesion of the coating system can arise if the adhesion promoter layer has not formed a sufficient bond with the substrate (adhesive bond), or if the immobilizing agent in the bulk of the coating system has not formed a sufficient bond with the adhesion promoter layer (cohesive bond). In the case of poor adhesion, this cross-cut test does not allow any conclusions to be drawn as to which of the two processes is responsible. Results: As shown in Figure 5, the samples (Sample 1_Slide No. 1 and Sample 1_Slide No. 2) of the coating system according to the invention have a favorable GT value of 1.Example 6 To ensure the adhesion strength and wetting behavior of the coating system according to the invention on a cleaned glass surface, the adhesion promoter layer (organosilane layer based on 3-(trimethoxysilyl)propyl methacrylate) was tested for its suitability in this regard. To achieve an optimal contact area between the coating and the glass surface, at which the adhesive interactions can be transferred, the adhesion promoter layer (primer layer) must, first and foremost, exhibit very good wetting behavior towards the immobilizing agent (LED-UV coating). The optimal wetting behavior of the adhesion promoter layer is determined in the test system by achieving: - a water contact angle of 55° ± 5° on the adhesion promoter layer; and - a background intensity of 40 ± 10 rfu. A contact angle measuring device from Krüss was used for the water contact angle measurements.Double-distilled water was used as the test liquid. To measure the static contact angle, a drop volume of 1-3 µl was continuously deposited onto the sample surface under investigation at a dosing rate of 2.0-4.0 µl / min. The contact angle analysis was performed automatically by the Krüss "ADVANCE" software. 05 / 12 / 2024 Without these parameters, a coating system regularly exhibits the effects of wetting disturbances, for example, in the form of crater formation in the immobilizing agent. For this reason, the relationship between the silane concentration of the adhesion promoter layer and the water contact angle was systematically investigated in order to determine optimal process conditions for applying the adhesion promoter layer to a cleaned substrate surface (silanization of a cleaned substrate surface).Furthermore, the formulation of the adhesion promoter layer is adapted to keep its intrinsic fluorescence low. Low intrinsic fluorescence of the adhesion promoter layer contributes to a low background intensity of the entire coating system and ensures that signals emitted by the immobilized, fluorescently labeled biomolecules can be detected without interfering background signals from the coating system itself. Since the low intrinsic fluorescence of the adhesion promoter layer is combined with the equally low intrinsic fluorescence of the immobilizing agent, the coating system according to the invention has a very low background intensity. Results: As shown in Figure 6, the desired water contact angle of 55 ± 5° is achieved with a silane concentration in a range of 0.005 to 0.01 mol / L and at a temperature of between 60 and 90°C.As further shown in Figure 7, adhesion promoter layers according to the invention achieve the required low background intensity of 40 ± 10 rfu. Example 7: PEG diacrylates and PEG methacrylates are produced via radical chain polymerization of, for example, poly(ethylene glycol) diacrylates (PEG), poly(ethylene glycol) methacrylates (PEG), and polythiols. These yield water-soluble, neutral polymers or highly water-swellable hydrogels. This ensures that an aqueous DNA solution can also penetrate the 3-dimensional polymer structure of the coating system and bind a larger number of biomolecules than would be possible with a polymer monolayer.Figure 8 schematically depicts a hydrogel with immobilized DNA probe molecules. It is intended to illustrate that such a coating system comprising a 3-dimensional (3D) polymer structure has a greater binding capacity for DNA probe molecules than a polymer monolayer (2D). Thus, using the coating system according to the invention, an amplification of the emitted fluorescence signal can be achieved after successful hybridization. Results: As shown in Figure 9, the coating system according to the invention with reactive epoxy groups for immobilizing biomolecules (left bar in each case) has a higher oligonucleotide binding capacity than a known 2D epoxy coating (reference; right bar in each case).Furthermore, it can be seen that the adsorption of the known 2D epoxy coating is close to saturation at a probe concentration of 5 µmol / L, whereas this is not yet the case with the 3D LED UV coating according to the invention, which, under the same conditions, allows for a further increase in adsorption. Example 8: In general, diagnostic coating systems are physically or chemically modified to ensure specific immobilization of biomolecules and thus precision in diagnostic results. As already shown in Example 2, certain embodiments of the coating system according to the invention contain HEMA to minimize nonspecific immobilization of biomolecules.Materials and Methods: - PBS buffer with a pH of 7.2 – pH 7.6 - PBS 0.05% Tween 20 - Skimmed milk powder solution - Goat serum labeled with the dye Dy547 (Goat Serum-NH2) To compare the non-specific binding of biomolecules to a coating system according to the invention, the following steps were carried out: - Glass substrates (slides) coated with the coating system were pre-scanned using a Tecan Scanner LS 400 before incubation in order to analytically determine the background intensity of the coating before the test. - After pre-scanning, the glass substrates coated with the coating system were blocked on a shaker at room temperature with an incubation time of 3 h using skimmed milk powder solution. 05 / 12 / 2024 - After blocking, Goat Serum-Dy547 was added to the incubation solution (and thus to the coated slides) and further incubated on a shaker at 4°C in the refrigerator.- After incubation, washing was carried out on the shaker. - To dry, the slides were centrifuged dry for 5 minutes at 1,300 rpm and RT. - After drying, the incubated slides were measured using a Tecan Scanner LS 400 to determine the signal intensity of the non-specifically bound Goat Serum-Dy547. As can be seen from Figures 10 and 11, the signal intensity of the immobilized Goat Serum-Dy547 on the inventive 3D LED UV epoxy coating with 451 or 425 rfu is approximately 68 to 71% lower compared to the 2D epoxy coating with 1487 rfu (Figure 10) or with 455 or 484 rfu compared to the 2D epoxy coating with 1490 rfu (Figure 11). Example 9: To adjust the contact angle of an aqueous sample on the inventive coating system, 2-hydroxyethyl-2-methacrylate was used as a hydrophilicity modulator in the immobilization agent.To compare the 3D epoxy coating system according to the invention with the known 2D epoxy coating (reference), the following test steps were carried out in parallel on both coatings: (1) Measurement of the intrinsic fluorescence; (2) Spotting of samples containing synthetically produced, single-stranded DNA fragments as biomolecules to be immobilized and an already fluorescently labeled DNA fragment as a positive control; (3) Immobilization of the biomolecules on the immobilization agent (30 minutes); (4) Measurement of the background fluorescence; (5) Hybridization with complementary CY3-labeled oligonucleotides in an automated microarray hybridization station (HS 4800 Tecan; approx. 2.5 hours) using the following washing solutions: 1. Washing solution: Pre Hyb 2. Washing solution: 2xSSC, 0.2% SDS 05 / 12 / 2024 3. Washing solution: 2xSSC 4. Washing solution: 0.2xSSC 5. Washing solution: deionized water 6.Washing solution: 0.1% SDS. Figure 12 shows that, as expected, a higher contact angle for water on a coated surface of a solid substrate results in an aqueous sample with a smaller spot diameter for the same volume. Figure 13 shows that, compared to the known 2D epoxy coating (reference), the coating system can not only absorb the higher sample density enabled by the increased contact angle over a smaller area, but can also permanently bind it, resulting in a significantly higher fluorescence intensity. The increased binding capacity of the coating system can be attributed to the 3-dimensional polymer structure of the immobilizing agent.Finally, it should be expressly pointed out that the above-described embodiments of the coating system according to the invention for immobilizing biomolecules, the solid-state substrate according to the invention for immobilizing biomolecules, the microarray or biochip according to the invention for immobilizing biomolecules, the functional solution according to the invention for producing an immobilizing agent and the method according to the invention for producing a coated solid-state substrate serve only to explain the claimed teaching, but do not limit it to the embodiments.

Claims

05 / 12 / 2024 Claims 1. A method for producing a coated solid substrate for immobilizing biomolecules, comprising the following steps: (a) applying an adhesion promoter layer to at least one surface of the solid substrate, wherein the contact angle of water on the adhesion promoter layer is preferably between 50° and 60°; (b) providing a functional solution comprising: ^ a crosslinking degree modulator comprising: acrylate monomers and / or acrylate oligomers with one acrylate ester group per molecule, preferably tetrahydrofurfuryl methacrylate monomers and / or oligomers; and / or 2-phenoxyethyl acrylate monomers and / or oligomers; and / or acrylate monomers and / or acrylate oligomers having more than one, preferably more than two or more than three acrylate ester groups per molecule, in particular comprising: trimethylolpropane ethoxy triacrylate monomers and / or oligomers;Dipentaerythritol penta- / hexacrylate monomers and / or oligomers; and / or functional dendrimers and / or dendrons, preferably highly branched bis(hydroxymethyl)propionic acid, acetylenic, highly branched bis(hydroxymethyl)propionic acid, azide and / or highly branched bis(hydroxymethyl)propionic acid, hydroxyl and / or highly branched poly(amidoamine) dendrimers and / or dendrons; ^ an immobilization modulator which provides reactive groups for the immobilization of biomolecules, the immobilization modulator preferably comprising: ester groups, preferably N-hydroxysuccinimide ester groups, glycidyl ester groups or isocyanate ester groups; ether groups, preferably glycidyl ether groups; epoxy groups; aldehyde groups; free carboxyl and / or free amino groups, as well as combinations thereof;and ^ a photoinitiator having an absorption maximum in the wavelength range between 315 and 400 nm, preferably a photoinitiator comprising: phenylglyoxylic acid methyl ester and diphenyl-(2,4,6-trimethylbenzoyl)phosphine oxide; ethylphenyl(2,4,6-trimethylbenzoyl)phosphinate; or phenylbis(2,4,6-trimethylbenzoyl)phosphine oxide; 05 / 12 / 2024 (c) applying the functional solution to the adhesion promoter layer; and (d) curing the functional solution using LED UV-A radiation in the wavelength range between 315 and 400 nm to form an immobilizing agent covalently bonded to the adhesion promoter layer, wherein the LED UV-A radiation causes a photochemically induced radical chain polymerization of the acrylate monomers and / or acrylate oligomers present in the functional solution and a 3-dimensional polymer structure of the immobilizing agent is formed by covalent crosslinking of the acrylate monomers and / or acrylate oligomers, in and on which reactive groups for immobilizing biomolecules are arranged.

2. The method according to claim 1, wherein the functional solution further comprises: ^ an additional raw material, preferably comprising hexanediol acrylate, poly(ethylene glycol) or polythiol; and / or ^ a hydrophilicity modulator, preferably 2-hydroxyethyl-2-methacrylate. 3.The method according to claim 2, wherein the functional solution comprises: ^ between 12 and 32 weight percent of the crosslinking degree modulator; ^ between 11 and 16 weight percent of the hydrophilicity modulator; ^ between 7 and 18 weight percent of the additional raw material; ^ between 20 and 50 weight percent of the immobilization modulator; and ^ between 3.5 and 6.5 weight percent of the photoinitiator.

4. The method according to any one of the preceding claims, wherein: ^ the application of the adhesion promoter layer to at least one surface of the solid substrate in step (a) is carried out by means of wet-chemical coating and / or chemical vapor deposition; and / or ^ the adhesion promoter layer predominantly comprises epoxysilane, (3-mercaptopropyl)trimethoxysilane, 3-(trimethoxysilyl)propyl methacrylate, polycyanurates, methacrylate phosphate esters or isocyanate-based surface additives; and / or. 05 / 12 / 2024 ^ the application of the functional solution to the adhesion promoter layer in step (c) is carried out by means of spin coating, slot die coating, airless spray application or ink jet processes; and / or ^ curing the functional solution in step (d) by means of LED UV-A radiation in the wavelength range from 340 to 390 nm or 350 to 390 nm or 350 to 390 nm or 360 to 390 nm or 370 to 390 nm or 380 to 390 nm or 385 nm or from 360 to 380 nm or 360 to 370 nm or from 365 nm.

5. The method according to any one of the preceding claims, wherein the solid-state substrate is a microarray or biochip for immobilizing biomolecules, preferably: nucleic acids, in particular deoxyribonucleic acids or ribonucleic acids; peptides; or proteins, in particular enzymes. 6.A coating system for immobilizing biomolecules on a surface provided with the coating system, comprising at least (a) an adhesion promoter layer and (b) an immobilizing agent covalently bonded to the adhesion promoter layer, characterized in that the immobilizing agent has a 3-dimensional polymer structure formed by covalent crosslinking of acrylate monomers and / or acrylate oligomers of a crosslinking degree modulator, wherein the 3-dimensional polymer structure is cured by a radical chain polymerization photochemically induced by UV-A radiation and contains reactive groups for immobilizing biomolecules. 7.The coating system according to claim 6, characterized in that: ^ at least some of the acrylate monomers and / or acrylate oligomers of the crosslinking degree modulator are: tetrahydrofurfuryl methacrylate monomers and / or oligomers; and / or 2-phenoxyethyl acrylate monomers and / or oligomers; or ^ at least some of the acrylate monomers and / or acrylate oligomers of the crosslinking degree modulator are more than one, preferably more. 05 / 12 / 2024 as two or more than three acrylate ester groups per molecule, in particular comprising: trimethylolpropane ethoxy triacrylate monomers and / or oligomers; dipentaerythritol penta- / hexacrylate monomers and / or oligomers; and / or ^ the immobilizing agent comprises highly branched functional dendrimers and / or dendrons, preferably highly branched bis(hydroxymethyl)propionic acid acetylenic, highly branched bis(hydroxymethyl)propionic acid azide and / or highly branched bis(hydroxymethyl)propionic acid hydroxyl and / or highly branched poly(amidoamine) dendrimers and / or dendrons.

8. The coating system according to claim 6 or claim 7, characterized in that: ^ the immobilizing agent comprises an additional raw material, preferably comprising hexanediol diacrylate, poly(ethylene glycol) or polythiol; and / or ^ the immobilizing agent comprises a hydrophilicity modulator, preferably 2-hydroxyethyl-2-methacrylate;and / or ^ the immobilization agent comprises residues of a starting radical of a photoinitiator, preferably of phenylglyoxylic acid methyl ester, of diphenyl-(2,4,6-trimethylbenzoyl)phosphine oxide, of ethyl (2,4,6-trimethylbenzoyl)phenylphosphinate, or of phenyl-bis(2,4,6-trimethylbenzoyl)phosphine oxide; and / or ^ the immobilization agent comprises an immobilization modulator which provides the reactive groups for immobilizing biomolecules, the immobilization modulator preferably comprising: ester groups, preferably N-hydroxysuccinimide ester groups, glycidyl ester groups or isocyanate ester groups; ether groups, preferably glycidyl ether groups; epoxy groups; aldehyde groups; free carboxyl and / or free amino groups, and combinations thereof.

9. The coating system according to any one of the preceding claims 6 to 8, wherein the 3-dimensional polymer structure of the immobilizing agent is formed by curing a functional solution comprising:;05 / 12 / 2024 ^ between 12 and 32 weight percent of the crosslinking degree modulator; ^ between 11 and 16 weight percent of the hydrophilicity modulator; ^ between 7 and 18 weight percent of the additional raw material; ^ between 20 and 50 weight percent of the immobilization modulator; and ^ between 3.5 and 6.5 weight percent of the photoinitiator.

10. The coating system according to any one of the preceding claims 6 to 9, wherein the UV-A radiation is an LED UV-A radiation in the wavelength range of 315 to 400 nm, preferably in the wavelength range of 340 to 390 nm or 350 to 390 nm or 350 to 390 nm or 360 to 390 nm or 370 to 390 nm or 380 to 390 nm or 385 nm or 360 to 380 nm or 360 to 370 nm or 365 nm.The coating system according to any one of the preceding claims 6 to 10, wherein the adhesion promoter layer predominantly comprises derivatives of epoxysilane, (3-mercaptopropyl)trimethoxysilane, 3-(trimethoxysilyl)propyl methacrylate, polycyanurates, methacrylate phosphate esters or isocyanate-based surface additives covalently bonded to acrylate monomers and / or acrylate oligomers of the immobilizing agent.

12. The coating system according to any one of the preceding claims 6 to 11, wherein the adhesion promoter layer can be applied to a surface by chemical vapor deposition, and / or wherein the immobilizing agent can be applied to the adhesion promoter layer by spin coating, slot die coating, airless spray application, or inkjet printing, and wherein the contact angle of water on the immobilizing agent is optionally greater than 60°, preferably between 60° and 70°. 13.The coating system according to one of the preceding claims 6 to 12, wherein the coating system has a solvent resistance to organic solvents, in particular to acetone, toluene or methanol, so that it does not show any detectable damage after treatment with organic solvents, in particular after treatment with toluene or methanol for 30 minutes each at 60 °C. 05 / 12 / 2024 14. A solid substrate for immobilizing biomolecules produced using a method according to any one of claims 1 to 5 and comprising at least one surface provided with a coating system according to any one of claims 6 to 13, wherein the substrate is preferably a glass substrate, plastic substrate, silicon substrate, or an oxide substrate, and is suitable for producing a microarray or a biochip.

15. The solid substrate according to claim 14, wherein the solid substrate is a microarray or biochip for immobilizing biomolecules, preferably for analyzing biomolecules contained in a sample, and wherein the biomolecules preferably comprise: nucleic acids, in particular deoxyribonucleic acids or ribonucleic acids; peptides;or proteins, in particular enzymes, and the microarray or biochip has a background intensity of less than 150 relative fluorescence units at an excitation wavelength of 532 nm.

16. Functional solution for use in a method according to one of claims 1 to 5 and / or for use in the production of an immobilizing agent of the coating system according to one of claims 6 to 13, at least comprising: ^ a crosslinking degree modulator comprising: acrylate monomers and / or acrylate oligomers with one acrylate ester group per molecule, preferably tetrahydrofurfuryl methacrylate monomers and / or oligomers; and / or 2-phenoxyethyl acrylate monomers and / or oligomers; and / or acrylate monomers and / or acrylate oligomers having more than one, preferably more than two or more than three acrylate ester groups per molecule, in particular comprising: trimethylolpropane ethoxy triacrylate monomers and / or oligomers;Dipentaerythritol penta- / hexacrylate monomers and / or oligomers; and / or functional dendrimers and / or dendrons, preferably highly branched bis(hydroxymethyl)propionic acid, acetylenic, highly branched bis(hydroxymethyl)propionic acid, azide and / or highly branched bis(hydroxymethyl)propionic acid, hydroxyl and / or highly branched poly(amidoamine) dendrimers and / or dendrons; 05 / 12 / 2024 ^ an immobilization modulator which provides reactive groups for the immobilization of biomolecules, the immobilization modulator preferably comprising: ester groups, preferably N-hydroxysuccinimide ester groups, glycidyl ester groups or isocyanate ester groups; ether groups, preferably glycidyl ether groups; epoxy groups; aldehyde groups; free carboxyl and / or free amino groups, and combinations thereof; and ^ a photoinitiator with an absorption maximum in the wavelength range between 315 and 400 nm, preferably a photoinitiator comprising: phenylglyoxylic acid methyl ester and diphenyl-(2,4,6-trimethylbenzoyl)phosphine oxide; ethylphenyl(2,4,6-trimethylbenzoyl)phosphinate; or phenylbis(2,4,6-trimethylbenzoyl)phosphine oxides. 17.Functional solution according to claim 16, further comprising: ^ an additional raw material, preferably comprising hexanediol diacrylate, poly(ethylene glycol), or polythiol; and / or ^ a hydrophilicity modulator, preferably 2-hydroxyethyl 2-methacrylate.

18. Functional solution according to claim 17, comprising: ^ between 12 and 32 weight percent of the crosslinking degree modulator; ^ between 11 and 16 weight percent of the hydrophilicity modulator; ^ between 7 and 18 weight percent of the additional raw material; ^ between 20 and 50 weight percent of the immobilization modulator; and ^ between 3.5 and 6.5 weight percent of the photoinitiator. 19.Use of a coating system according to one of claims 6 to 13 for immobilizing biomolecules on a surface of a microarray or a biochip provided with the coating system, wherein the biomolecules are preferably: nucleic acids, in particular deoxyribonucleic acids or ribonucleic acids; peptides; or proteins, in particular enzymes. 05 / 12 / 2024 20. A microarray or biochip for immobilizing biomolecules, preferably for analyzing biomolecules contained in a sample, comprising the coating system according to any one of claims 6 to 13 and / or the solid-state substrate according to claim 14 or 15, wherein the biomolecules are preferably: nucleic acids, in particular deoxyribonucleic acids (DNA) or ribonucleic acids (RNA); peptides; or proteins, in particular enzymes.

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