Measurement device for analysing the optical response of diffractive biosensitive materials

Abstract

The invention relates to a measurement device for analysing the optical response of diffractive biosensitive materials. Said device comprises a diffractive biosensitive material; a substrate on which the diffractive biosensitive material is supported; a light source that irradiates the diffractive biosensitive material, so that said material generates the diffractive optical response that allows the amount of target compound present in the sample to be determined, and wherein the light source directs its radiation towards the diffractive biosensitive material at an angle of incidence; a transmission area and a reflection area in which the diffractive optical response takes place; a fluidic system for placing samples in contact with the diffractive biosensitive material; photodetectors, which are arranged to record the intensity of the incident radiation and / or the intensity of the diffractive optical response; and light blocking elements around the photodiodes.

Description

[0001] DESCRIPTION

[0002] MEASURING DEVICE FOR THE ANALYSIS OF THE OPTICAL RESPONSE OF DIFFRACTIVE BIOSENSITIVE MATERIALS

[0003] TECHNICAL SECTOR

[0004] The invention relates to a handheld device for recording optical signals generated by diffractive biosensitive materials as a result of biorecognition assays. To this end, the proposed device consists of a set of elements arranged to isolate and quantify the diffractive optical signals, determine the result of the analysis, and communicate it to the user.

[0005] BACKGROUND OF THE INVENTION

[0006] It is well known that chemical and biological analysis systems are of great importance in the commercial and social spheres, as they are used in products that improve society's quality of life. These technologies play a central role in areas such as disease detection and prevention, contaminant determination, and industrial quality control. Innovation in the field of chemical and biological analysis systems is essential both to improve people's quality of life and health, and to drive commercial and industrial development through the creation of new products and technologies that meet market needs and demands.

[0007] In this sense, by way of example, various measurement systems are known for recording optical signals generated by diffractive biosensors, among which the following stand out:

[0008] Optical table mounted systems, typically employed for proof of concept studies in research environments involving applied optics [1-8].

[0009] The adaptation of compact disc technology, with discs modified to act as supports for periodic nanostructures of bioreceptors, and disc readers modified to scan their diffractive response

[0010] .

[0010] Table-top analysis equipment based on periodic nanostructures of bioreceptors, which are complex and bulky, and deviate from the Point-of-care guidelines [7,8,11 , 12],

[0011] References:

[0012] [1] Analytica Chimica Acta (2018), 1033, 173-179. https: / / doi.Org / 10.1016 / j.aca.2O18.05.060

[0013] [2] Biosensors and Bioelectronics (2021), 176, 112916. https: / / doi.Org / 10.1016 / j. bios.2020.112916

[0014] [3] ACS Applied Materials and Interfaces (2022),

[0015] 14, 41640-41648. https: / / doi.org / 10.1021 / acsami.2c12808

[0016] [4] Biosensors (2023), 13, 214. https: / / doi.Org / 10.3390 / bios13020214

[0017] [5] Biosensors and Bioelectronics (2021 ) 193, 113561. https: / / doi.Org / 10.1016 / j . bios.2021 .1 13561

[0018] [6] Biosensors (2023) 13, 312. https: / / doi.org / 10.3390 / bios13030312

[0019] [7] Nature Nanotechnology (2017) 12, 1089-1095. https: / / doi.Org / 10.1038 / NNAN0.2017.168

[0020] [8] Sensors and Actuators B: Chemical

[0021] (2021 ) 349, 130746. https: / / doi.Org / 10.1016 / j.snb.2021.130746

[0022] [9] Nature Microbiology (2019) 4, 46-54. https: / / doi.org / 10.1038 / s41564- 018-0295-3

[0023]

[0010] Analytical Chemistry (2017), 89, 9002-9008. https: / / doi.Org / 10.1021 / acs.analchem.7b01649

[0024]

[0011] PLOS Neglected Tropical Diseases

[0025] (2014), 8, 3002. https: / / doi.org / 10.1371 / journal.pntd.0003002

[0026]

[0012] US 2022 / 0298555 A1

[0027] It is well known that an innovative and advantageous strategy for developing chemical and biological analysis technologies consists of combining biosensors with light diffraction techniques. Among other aspects, the integration of macromolecules such as antibodies, enzymes, or nucleic acids allows for the development of selective and sensitive biosensors. Furthermore, diffractive techniques are well known for their great utility in detecting target compounds without labeling, in real time, and solving significant problems of nonspecific adsorption in the direct analysis of untreated biological samples [1-6].

[0028] This combination of biosensing and diffraction is based on materials structured at the micrometer and / or nanometer scale, which diffract incident light and integrate biomacromolecules responsible for specific biorecognition. When these materials come into contact with the sample to be analyzed, the macromolecules they comprise interact with the target compounds present in the sample, and this phenomenon generates a measurable change in their diffractive response. This change can determine the presence and / or concentration of the target compound.

[0029] In the state of the art there are two methods to address this strategy: through periodic nano / micro-structures of bioreceptors fabricated on solid substrates [1-4, 7, 8], and through bioreceptors included in three-dimensional matrices structured by holographic means [5-6].

[0030] A fundamental aspect of chemical and biological analysis technologies is that they should be cheap, simple, compact, robust, portable, and easy to use by non-specialized users, as pointed out by the World Health Organization [9]. With the prevalence of infectious and chronic diseases, the increase in the geriatric population, and the new post-pandemic dynamics, there is a great demand for analytical systems with these characteristics, which is accompanied by a high social and economic impact. Compared to standard chemical and biological analysis technologies, biosensor devices with these characteristics allow the analysis to be carried out and the result to be obtained in the same place and at the same time as it is needed, and by non-specialized users for the performance and interpretation of the test. These types of devices, as previously mentioned, are defined in the bioanalytical field as Point-of-care or Point-of-need systems.

[0031] Compared to these systems described and known in the state of the art, the technology object of the present invention consists of a handheld device that is small, portable, compact, and easy to use by non-specialized users. These features allow the advantages of diffractive biosensors to be implemented in point-of-care scenarios. That is, it allows the direct analysis of complex, unprocessed biological samples without labeling, at the same time and place where the result is required, and by non-specialized users.

[0032] In this regard, we know what was disclosed in document US2019017938A1, which refers to a diffractive biosensor for the detection of biomolecules.

[0033] Also known is that described in document WO2021170601 A1, which discloses a portable diffractive analysis device for analyzing molecular interactions.

[0034] It is known what was disclosed in document W020200015872A1 where a diffractive biosensor for the selective detection of molecules in a grid with biological material is described.

[0035] These three documents improve on the previously mentioned technology; however, compared to the present invention, they present significant differences in materials and detection principles. The devices described in those three documents are based on waveguides, while the present invention is based on free-space diffraction.

[0036] Going into greater detail, in the three aforementioned patents, the diffractive biosensitive material is arranged on a waveguide, which in most cases is represented as a planar waveguide (slab type). In these cases, the diffractive biosensitive material interacts with the evanescent field of the radiation guided by the waveguide. These systems necessarily require coupling and / or decoupling of the light in the waveguides.

[0037] Compared to these technologies, the present invention is not based on waveguides; rather, the radiation propagates unguided (in free space) and thus interacts with the diffractive biosensitive material. This difference is not trivial from a technical perspective; it entails significant differences in the implementation of the devices and offers significant advantages over what is disclosed in those patents. It should be noted that waveguides are nanostructured materials, which typically involve complex and expensive raw materials and manufacturing methods. The present invention does not require waveguides, so compared to the devices proposed in the three patents, it offers significant advantages in terms of simplicity, handling versatility, and cost.

[0038] It should also be noted that the ability to analyze biological samples in the medical field is essential for this industrial sector. In this sector, it is often a requirement that materials be disposable and non-reusable. In this sense, using simple and inexpensive substrates like the substrates proposed in the present invention, instead of more expensive materials like the waveguides proposed in the three previous patents, represents an advantage in terms of the possibility of materializing disposable systems. Furthermore, compared to disposing of waveguides, disposing of simple substrates offers advantages in terms of environmental and economic sustainability.

[0039] Additionally, light coupling and / or decoupling in waveguides tends to be a less robust process and requires additional optical elements, which complicate the final device and restrict its applicability. The present invention does not involve coupling / decoupling; it simply irradiates in free space. Therefore, compared to the three patents, it offers advantages in terms of the robustness and simplicity of the final device. This fact also positively impacts the sensitivity of the resulting analyses.

[0040] Finally, in the devices suggested in these three patents, the diffractive optical response is concentrated in a microscopic spot, which requires additional optical elements (lenses) to focus and quantify the diffractive optical response. In the present invention, the diffractive optical response is not focused and can be detected directly with conventional photodetectors. Compared to the three patents, this provides simplicity and reduces costs in the final device.

[0041] Taking these aspects into account, the applicant is not aware of any measuring device for the analysis of the optical response of diffracted biosensitive materials as effective as the one described below. EXPLANATION OF THE INVENTION

[0042] The invention consists of a measuring device for the analysis of the optical response of biosensitive materials, which is compact and portable, and which has the particularity of comprising the following components:

[0043] A diffractive biosensitive material. This material contains receptors (antibodies, enzymes, nucleic acids, or others) that are capable of interacting with target compounds present in a sample. When a sample containing a target compound comes into contact with the biosensitive material, the bioreceptor within the biosensitive material interacts with the target compound, and this interaction changes the diffractive response of the material. Examples of diffractive biosensitive materials include layers of bioreceptors immobilized on solid substrates and structured into a diffraction grating pattern, or hydrogel-like polymer matrices incorporating bioreceptors and containing a diffractive pattern etched into the core or surface of the material.

[0044] A substrate or support on which the diffractive biosensitive material is mounted. This material may be transparent, reflective, partially reflective, or contain both reflective and transparent areas. The support may also be opaque or contain opaque zones to adjust the optical response. Some examples of materials that can be used as substrates are glass, synthetic plastics (polystyrene, polycarbonate, cyclic polyolefins, polypropylene, or others), metals (steel, gold, aluminum, or others), fibers and membranes (cellulose, nitrocellulose, or others), silicon and silicon derivatives, as well as the previous examples with physicochemical modifications to adjust the optical and chemical properties of the substrate. Likewise, the substrate may not be included in diffractive biosensitive materials that do not require it.A single substrate may also include several portions of different diffractive biosensitive material, to perform more than one chemical and / or biological analysis.

[0045] A light source irradiates the diffractive biosensitive material, so that it generates the diffractive optical response that allows the quantity of target compound present in the sample to be determined. Some examples of light sources suitable for this invention are laser systems (pointers, heads, or others), LED and OLED sources, or polychromatic lamps (incandescent, halogen, fluorescent, or others). The light source directs its radiation towards the diffractive biosensitive material at an angle of incidence where 0 e < <90 e , that is, it can range from 0 degrees (perpendicular irradiation) to 90 degrees.

[0046] A transmission zone and a reflection zone in which the diffractive optical response takes place, defined according to the position of the diffractive biosensitive material with respect to the light source.

[0047] A fluidic system for bringing samples into contact with the diffractive biosensitive material. Some examples of fluidic systems include incubation chambers, channels, and others.

[0048] At least one photodetector, which is arranged to record the intensity of the incident radiation and / or the intensity of the diffractive optical response in the reflection zone and / or in the transmission zone. It may integrate at least one which may be in the reflection zone or in the transmission zone. Some examples of photodetectors suitable for this invention are photodiodes, photomultipliers, phototransistors, photovoltaic cells, CCD (Charge-Coupled Device) cameras, CMOS (Complementary Metal-Oxide-Semiconductor) cameras, silicon image sensors (SIS), and infrared (IR) cameras.

[0049] Light-blocking elements used to isolate the diffractive optical response and eliminate potential optical interference. They are physical barriers made of opaque materials placed around the photodiodes and / or the diffracted optical response.

[0050] An identification code located on the support, which allows for automatic identification of the type of analysis using a compatible detector / receiver. Some examples of identification codes and detectors suitable for this research include barcodes in conjunction with barcode readers, REID tags in conjunction with REID readers, or QR codes in conjunction with cameras on smart devices (mobile phones, tablets, computers, or similar). Identification codes and / or their readers may also not be included on the device.

[0051] An electronic controller controls the system's components and computes the response of biological and chemical assays. This controller may include elements such as a CPU, an analog-to-digital converter, memory, transistors, resistors, input and output connections, and others. The controller connects to the various components of the device via electrical signal transmission cables.

[0052] A battery or power source that powers the electronic controller, light source, detectors, and other components of the device. Examples of batteries compatible with this device include lithium-ion batteries, lithium-ion polymer batteries, nickel-metal hydride batteries, nickel-cadmium batteries, alkaline batteries, and zinc-carbon batteries.

[0053] A housing that incorporates all the system elements within or on its surface. The housing may include holes or openings for introducing the diffractive biosensitive material inside and recording its optical response. The housing is made of a polymeric material (polycarbonate, polystyrene, polypropylene, acrylonitrile butadiene styrene, polylactic acid, or others), metallic material (steel, aluminum, titanium, or others), or a combination of the above.

[0054] Switches or buttons to manually operate and control the device and perform chemical and biological analysis measurements.

[0055] A display serves as an interface for communicating data (instructions, results, etc.) to the device's user. Examples of suitable displays for this device include liquid crystal displays, LED displays, or touchscreens. Alternatively, the electronic controller may include a wireless technology port (Bluetooth, NFC, Wi-Fi, or others) that transfers data to an external device (mobile phone, tablet, computer, smartwatch, or others), and the display may be omitted.With all these elements, as previously mentioned, the present invention, compared to known technologies, is not based on waveguides, but on the radiation propagating in an unguided manner (free space) and thus interacting with the diffractive biosensitive material, and it is considered that this difference is not trivial from a technical point of view, since it entails significant differences in the materialization of the devices, and provides significant advantages compared to those other known technologies.

[0056] Going into other types of details, in possible embodiments of the invention, the device and the housing have dimensions between 5 and 30 cm long, between 3 and 20 cm high, and between 3 and 30 cm wide, and the housing is composed of a synthetic polymer.

[0057] In a possible embodiment of the invention, the diffractive biosensitive material consists of a biolayer of bioreceptors (proteins and nucleic acids) immobilized on a glass substrate according to a one-dimensional periodic pattern.

[0058] In another embodiment of the invention, the diffractive biosensitive material consists of a hydrogel-type matrix with bioreceptors (proteins or nucleic acids) in its composition, and in which a periodic network has been engraved in its volume or on its surface by interference of two lasers or by replication of a master with a surface relief network.

[0059] In another embodiment of the invention, the diffractive biosensitive material consists of a solid support composed of a synthetic polymer (polystyrene, polycarbonate, etc.) or a silicon-based material (silicon, glass, silicon nitride, etc.), which contains a periodic network on its surface on which a layer of bioreceptors (proteins or nucleic acids) is arranged.

[0060] In a possible embodiment of the invention, the light source is a laser with a wavelength between 400 nm and 1000 nm, and the photodetectors are photodiodes located in the transmission zone.

[0061] In another possible embodiment of the invention, the light source is a halogen lamp or an LED, and the photodetectors are CCD cameras located in the reflection zone. In one possible embodiment of the invention, the housing has a side opening through which a support with diffractive biosensitive material irradiated perpendicularly (0 degrees) is inserted.

[0062] In another possible embodiment of the invention, the housing has a lateral opening aligned to introduce a support with diffractive biosensitive material to be irradiated obliquely (between 0 and 90 degrees).

[0063] In a possible embodiment of the invention, the support includes a barcode, the device includes a barcode reader, and the housing includes an LCD screen that acts as an interface for the user.

[0064] In another possible embodiment of the invention, the support includes a QR code, and the electronic controller includes a Bluetooth port to transmit information regarding the analysis to the user through a smart device.

[0065] It should be noted that throughout the description and claims, the term “comprises” and its variants are not intended to exclude other technical features or additional elements.

[0066] BRIEF DESCRIPTION OF THE DRAWINGS

[0067] To complement the description being made and in order to help better understand the characteristics of the invention, a set of drawings is attached as an integral part of said description, in which the following has been represented for illustrative and non-limiting purposes:

[0068] Figure 1 shows a schematic representation of a measuring device that is the object of the present invention and the elements that comprise it.

[0069] Figure 2 shows in detail the scheme where diffractive biosensitive material, the substrate and the fluidic system.

[0070] Figure 3 shows an external perspective view of a possible embodiment of the device object of the present invention, specifically where the substrate with the diffractive biosensitive material is introduced laterally into the device to be irradiated perpendicularly.

[0071] DESCRIPTION OF AN EMBODIMENT OF THE INVENTION

[0072] As can be seen in the set of drawings, an embodiment of the measuring device for the analysis of the optical response of diffractive biosensitive materials comprises a housing (17) that houses all the components of the device and that comprises holes or openings (18) to introduce a diffractive biosensitive material (1) inside and record its optical response; which is a handheld device, small, portable, compact and easy to use by non-specialized users; and has the particularity of comprising: a diffractive biosensitive material (1), which contains receptors that are capable of interacting with target compounds present in a sample; a substrate (2) on which the diffractive biosensitive material is supported; a light source (3) that irradiates the diffractive biosensitive material, so that it generates the diffractive optical response (4) that allows determining the quantity of target compound present in the sample;and where the light source directs its radiation towards the diffractive biosensitive material at an angle of incidence (5), where 0; e < <90 e ; a transmission zone (6) and a reflection zone (7) in which the diffractive optical response takes place, defined according to the position of the diffractive biosensitive material with respect to the light source. a fluidic system (8) for bringing samples into contact with the diffractive biosensitive material; photodetectors (9), which are arranged to record the intensity of the incident radiation (10) and / or the intensity of the diffractive optical response in the reflection zone and / or in the transmission zone; light blocking elements (11) used to isolate the diffractive optical response and eliminate possible optical interferences, which are physical barriers of opaque materials that are placed around the photodiodes and / or the diffracted optical response.

[0073] In addition to those essential elements for solving the technical problem posed in the present invention, this embodiment may have an identification code (12) located on the substrate, which allows the type of analysis to be automatically identified using a compatible detector / receiver (13). It may have an electronic controller (14) to control the elements of the system and compute the response of the biological and chemical tests. It may have a battery (16) or power supply that feeds the electronic controller, the light source, the detectors, and other elements that make up the device; the housing (17) may have switches or buttons (19) to manually operate and control the device and carry out the measurements of the chemical and biological analyses; and it may have a screen (20) as an interface to communicate data.

Claims

CLAIMS 1 - Measuring device for the analysis of the optical response of diffractive biosensitive materials, comprising a housing (17) that houses all the components of the device and comprising holes or openings (18) to introduce a diffractive biosensitive material (1) inside and record its optical response, characterized in that it comprises: a diffractive biosensitive material (1), which contains receptors capable of interacting with target compounds present in a sample; a substrate (2) or support on which the diffractive biosensitive material is supported; a light source (3) that irradiates the diffractive biosensitive material, so that it generates the diffractive optical response (4) that allows determining the quantity of target compound present in the sample; and where the light source directs its radiation towards the diffractive biosensitive material at an angle of incidence (5) that, where 0 e < <90 e. a transmission zone (6) and a reflection zone (7) in which the diffractive optical response takes place, defined according to the position of the diffractive biosensitive material with respect to the light source; a fluidic system (8) for bringing samples into contact with the diffractive biosensitive material; photodetectors (9), which are arranged to record the intensity of the incident radiation (10) and / or the intensity of the diffractive optical response in the reflection zone and / or in the transmission zone; and light-blocking elements (11) for isolating the diffractive optical response and eliminating possible optical interference, which are physical barriers made of opaque materials that are placed around the photodiodes. 2- A device, according to claim 1, comprising an identification code (12) located on the automatic identification support of the type of analysis using a compatible detector / receiver (13). 3.- A device according to claim 1, comprising an electronic controller (14) for controlling the elements of the device and computing the response of the biological and chemical tests. 4.- A device according to claim 3, wherein the electronic controller (14) includes a Bluetooth port for transmitting information to the user through a smart device.

5. A device according to claim 1, comprising a battery (16) or power supply that powers the components of the device. 6.- A device, according to claim 1, comprising switches or buttons (19) for manual operation and control of the device. 7.- A device according to claim 1, comprising a display (20) as an interface for communicating data. 8.- A device according to claim 1, wherein the diffractive biosensitive material (1) consists of a biolayer of bioreceptors, such as proteins and nucleic acids, immobilized on a glass substrate according to a one-dimensional periodic pattern. 9.- A device according to claim 1, wherein the diffractive biosensitive material (1) consists of a hydrogel-type matrix with bioreceptors, such as proteins or nucleic acids, in its composition, and in which a periodic network is recorded in its volume or on its surface by interference of two lasers or by replication of a master with a surface relief network. 10.- A device according to claim 1, wherein the diffractive biosensitive material (1) consists of a solid support composed of a synthetic polymer or a silicon-based material, which contains a periodic network on its surface on which a layer of bioreceptors, such as proteins or nucleic acids, is arranged. 1.- A device according to claim 1, wherein the photodetectors (9) are photodiodes located in the transmission zone. 12.- A device according to claim 1, wherein the photodetectors (9) are CCD cameras located in the reflection zone. 13.- A device according to claim 1, wherein the light source is a halogen lamp or an LED. 14.- A device according to claim 1, wherein the support includes a QR code. 15.- A device according to claim 1, wherein the light source (3) is a laser with a wavelength between 400 nm and 1000 nm.

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