Device for integrated treatment of biological sample

Abstract

The invention relates to a device and a corresponding method for the integrated processing of biological samples by means of centrifugal forces applied along the vertical axis of the device. The device comprises at least a first and a second compartment for performing successive steps of sample processing, the two compartments being vertically arranged and in fluid communication with each other, where each compartment comprises a microfluidic structure at its respective distal end. The capillaries of the first microfluidic structure have a larger average diameter than the capillaries of the second microfluidic structure, enabling a biological sample to pass through the microfluidic structures by increasing centrifugal force along the vertical axis of the device when traveling from the first compartment to the second compartment of the device.

Description

Technical Field

[0001] The present invention relates to an apparatus and a corresponding method for the integrated processing of biological samples by means of a centrifugal force applied only along the vertical axis of the apparatus, wherein the centrifugal force increases as the sample processing progresses. Background Art

[0002] In the past decade, the development of integrated on-site or point-of-care devices for biomolecular diagnostics has been driven by the miniaturization and automation of the analytical protocols to be employed. Many such devices are based on microfluidic systems. The obvious advantages of microfluidics include reduced consumption of samples and reagents, increased efficiency, and relatively fast reaction times.

[0003] However, there are still some obstacles to overcome in order to further improve automation. In many cases, biomolecular analysis is not hindered by the actual diagnostic test itself, but rather by the handling of biological samples during the period from collection of the biological sample to introduction into the test device. Biological samples typically need to undergo some form of purification, fractionation, filtration, dilution, concentration, digestion, or preservation in order to be suitable for downstream analysis (e.g., not to clog the device due to high viscosity), thereby providing reliable and accurate results.

[0004] These preparatory steps of sample handling usually require long and tedious manual handling and extensive use of disposable consumables. Different devices are often required to perform different handling steps, thereby increasing the inherent error of the analysis due to unnecessary sample manipulation. The protocols employed are usually not standardized, which interferes with reproducibility and quality control, and interferes with the setup of automated workflows and sample handling in high-throughput programs.

[0005] Therefore, there is a continuing need for improved devices and corresponding methods for handling biological samples, particularly with regard to complex diagnostic applications such as genomics analysis, proteomics analysis, and metabolomics analysis. In particular, there is a need for a single and easy-to-use microfluidic device that integrates multiple steps of sample handling and does not require trained personnel to operate.

[0006] Therefore, an object of the present invention is to provide such a device for the integrated processing of biological samples and its corresponding application method. Summary of the Invention

[0007] In one aspect, the present invention relates to a device for processing a biological sample by means of a centrifugal force applied along a vertical axis of the device, the device comprising: (i) at least a first compartment for performing a first step of sample processing, the first compartment comprising a structure for loading a biological sample at its proximal part and a first microfluidic structure at its distal part; and (ii) at least a second compartment for performing a second step of sample processing, a proximal end of the second compartment being in fluid communication with a distal end of the first compartment, the second compartment comprising a second microfluidic structure at its distal part, the second step of sample processing being consecutive to the first step of sample processing; wherein the first microfluidic structure comprises a capillary having a first average diameter, the second microfluidic structure comprises a capillary having a second average diameter, the first average diameter being greater than the second average diameter, such that a biological sample can be made to pass through the first microfluidic structure by applying a first centrifugal force along the vertical axis of the device, and such that a biological sample can be made to pass through the second microfluidic structure by applying a second centrifugal force along the vertical axis of the device, the second centrifugal force being greater than the first centrifugal force.

[0008] In a preferred embodiment, the first microfluidic structure exhibits a more hydrophilic property than the second microfluidic structure. To this end, the material for manufacturing the first microfluidic structure may exhibit a more hydrophilic property than the material for manufacturing the second microfluidic structure; and / or the first microfluidic structure may have a functional surface modification so as to exhibit a more hydrophilic property than the second microfluidic structure.

[0009] In other specific embodiments, the processing of the biological sample includes at least partial purification of the sample. In a preferred embodiment, the processing of the biological sample includes at least two steps selected from the group consisting of: collection of the sample, filtration of the sample, desalting of the sample, concentration of the sample, dilution of the sample, solubilization and / or digestion of the sample, depletion of a part contained in the sample, and enrichment of a part contained in the sample.

[0010] In a particular embodiment, the device is integrally formed as a whole. Alternatively, at least one compartment of the device is formed as a separate component, which can be connected to the remaining one or more compartments of the device.

[0011] In a further particular embodiment, the device is configured to be placed in a reaction vessel and used together with the reaction vessel, wherein the volume of the reaction vessel is from 0.1 ml to 50 ml, preferably from 0.2 ml to 10 ml, more preferably from 0.5 ml to 2.5 ml.

[0012] In other specific embodiments, the device further includes a compartment for sample loading, wherein a distal end of the compartment for sample loading may be connected to a proximal end of the at least first compartment such that the two compartments are in fluid communication.

[0013] In still some other specific embodiments, the device further includes a reagent reservoir, wherein a distal end of the reagent reservoir may be connected to a proximal end of the compartment for sample loading such that the two compartments are in fluid communication; or a distal end of the reagent reservoir may be connected to a proximal end of the at least first compartment such that the two compartments are in fluid communication, particularly wherein the reagent reservoir is pre-filled with a reagent.

[0014] In still some other specific embodiments, the device further includes a sample collection reservoir for collecting the processed sample, wherein a proximal end of the sample collection reservoir may be connected to a distal end of the at least second compartment such that the two compartments are in fluid communication.

[0015] In another aspect, the present invention relates to a method for processing a biological sample by means of centrifugal force applied along a vertical axis of a processing device, the method comprising: (i) introducing a biological sample into the device as defined above; (iii) performing a first step of sample processing in at least a first compartment of the device by applying a first centrifugal force along the vertical axis of the device such that the sample can pass through a first microfluidic structure; and (iii) performing a second step of sample processing in at least a second compartment of the device by applying a second centrifugal force along the vertical axis of the device such that the sample can pass through a second microfluidic structure, wherein the second centrifugal force is greater than the first centrifugal force.

[0016] In specific embodiments, the method further includes: loading a biological sample into a compartment for sample loading connected to at least a first compartment of the device; and / or releasing a reagent from a reagent reservoir connected to the compartment for sample loading of the device or connected to at least a first compartment after the biological sample has been introduced into the device.

[0017] In other specific embodiments, the method further includes: collecting the processed sample in a sample collection reservoir connected to at least a second compartment of the device.

[0018] In a preferred embodiment, the biological sample is selected from the group consisting of blood, plasma, serum, saliva, urine, nasopharyngeal swab, oropharyngeal swab, and tissue sample.

[0019] In another aspect, the invention relates to the use of the device as defined above for processing biological samples to be subjected to "omics" applications, in particular where said "omics" applications are selected from the group consisting of genomics, transcriptomics, proteomics, lipidomics, glycomics, microbiomics and metabolomics. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 :(A) Photograph of an exemplary device according to the invention. The device is configured to be used (or configured to have the shape and dimensions of) a standard 2 ml reaction tube during centrifugation (shown in the upper row). The device includes a processing unit having two compartments (third from the left in the lower row) configured according to the intended application. The processing unit has an interface for sample uptake at its proximal end. After sample uptake, a reagent reservoir (second from the left in the lower row) is connected to the processing unit. After loading the required reagents, the reagent reservoir is closed with a screw cap (left side in the lower row). After processing the purified sample by applying a centrifugal force along the vertical axis of the device, the processed sample is collected in a sample collection reservoir (right side in the lower row) connected to the distal end of the processing unit. (B) Photograph of the exemplary device in assembled form.

[0021] Figure 2 : Diagram of the general principle on which the device according to the invention is based. The device includes at least two compartments for sample processing, each compartment including a microfluidic structure. The average diameter of the capillaries of the first microfluidic structure (or capillary barrier) is greater than the diameter of the capillaries of the second microfluidic structure, so that an increased centrifugal force is required for the sample to cross the capillary barrier when traveling from the first compartment to the second compartment. The device is configured such that it can be used with a standard reaction tube (here a 2 ml reaction tube).

[0022] Figure 3 : Diagram of an exemplary device according to the invention suitable for processing urine samples. The device includes two compartments, which are respectively used for desalting of urine samples and removal of a large amount of proteins. For this purpose, the first compartment is equipped with a filter membrane for desalting, and the second compartment is equipped with an affinity matrix for binding a large amount of proteins. Each compartment has a microfluidic structure (capillary barrier) at its distal end, wherein the first microfluidic structure has more capillaries with a larger average diameter compared to the second microfluidic structure. The processed sample is collected in a sample collection reservoir. The device is configured such that it can be used with a standard 50 ml reaction tube.

[0023] Figure 4: Diagram of an exemplary multi-component device according to the present invention. The device includes a reagent reservoir (A) having a closed bottom surface (e.g., in the form of a membrane), the closed bottom surface having a rupture point that is ruptured by a push column when the reservoir is threadedly connected to a central processing unit (B). The processing unit includes at least two compartments for sample processing and is threadedly connected to a sample collection reservoir (C). The assembled device is shown in (D).

[0024] Figure 5 : Diagram (A) of an exemplary workflow for sample processing according to the present invention. A biological sample is applied to the first compartment of the device placed in a 2 ml standard reaction tube using a pipette. A reagent reservoir containing a buffer and other reagents for performing sample processing is connected to the first compartment of the device. The reaction tube is closed, and sample processing is performed by applying a centrifugal force in a microcentrifuge. The device has an open bottom surface such that the processed sample is collected in the reaction tube and is ready for further analysis after the device is removed from the tube. (B) The corresponding device employed is configured to fit in a standard reaction tube. The portion shown in light gray includes the first compartment and the second compartment, while the portion shown in dark gray represents the reagent reservoir connected to the first compartment. A cross-sectional view (C) of the corresponding device shown in (B) shows two microfluidic structures at the distal portions of the first compartment and the second compartment, respectively. The bottom surface of the device is open such that the processed sample can be collected in the reaction tube.

[0025] Figure 6 : Diagram of the effect of a polymer material used to fabricate a microfluidic structure on the following centrifugal forces (in revolutions per minute (rpm)): the centrifugal force required to completely transfer water through a capillary of a certain average diameter (in μm, shown in the range between 50 μm and 1.1 mm). An exemplary analysis of a microfluidic structure made of polypropylene (A) or polytetrafluoroethylene (B) is shown. The thickness of the microfluidic structure employed in the analysis is 1 mm. Centrifugation is performed at a temperature of 20 °C using a 2 ml reaction tube with a rotor having a diameter of 24 cm.

[0026] Figure 7: Graph showing the influence of the advancing contact angle (shown in the range between 90° and 150°) on the following centrifugal forces (in revolutions per minute (rpm)): the centrifugal forces required to completely transfer water through capillaries (made of an arbitrary polymer) with an average diameter of 100 μm (upper curve) and 1 mm (lower curve). The advancing contact angle is the highest possible contact angle of a liquid droplet located on the surface. The advancing contact angle is measured when a liquid (here water) wets a previously dry surface by gradually applying the liquid to the surface with a needle. The angle can be measured when (or just as) the contact line starts to move. The thickness of the microfluidic structure employed in the analysis is 1 mm. Centrifugation is carried out at a temperature of 20 °C using a 2 ml reaction tube with a rotor having a diameter of 24 cm. Detailed Description

[0027] The present invention is based on the unexpected discovery that the processing and preparation of biological samples for downstream applications can be performed solely by means of vertical centrifugal force in a single device, thereby providing a significantly more labor-saving, simplified, and cost-effective process. The hands-on time for loading the sample and placing the device in the centrifuge is limited to just a few minutes. Depending on the applied processing protocol, the hands-on time and the total cost per trial can be reduced by at least 30% to 60%.

[0028] The device has a special design, with a vertical arrangement of at least two compartments that are in fluid communication with each other and having microfluidic structures at their respective distal ends. Each compartment is adapted to perform one step of sample processing, where the transport from one compartment to the successive compartment is controlled by the applied centrifugal force. A biological sample is introduced into the device and the biological sample is transported by applying centrifugal force along the vertical axis of the device, where the resistance of the sample passing through the first microfluidic structure is lower than the resistance of passing through the second microfluidic structure. This is achieved by the configuration of the microfluidic structures, where the capillaries of the first microfluidic structure in particular have a larger average diameter than the capillaries of the second microfluidic structure. Optionally, the first microfluidic structure also exhibits a more hydrophilic nature than the second microfluidic structure. Thus, the processing of the biological sample proceeds unidirectionally along the vertical axis of the device.

[0029] The present invention will be described hereinafter with reference to specific embodiments and certain figures, but the present invention should be understood as not being limited thereto, but only being defined by the appended claims. The present invention can be suitably implemented in the absence of any element or limitation not specifically disclosed herein.

[0030] When the term "comprising" is used herein, it does not exclude other elements or steps. For the purposes of the present invention, the term "consisting of" is considered a preferred embodiment of the term "comprising". If a group is defined hereinafter as including at least a certain number of embodiments, this should also be understood as disclosing a group consisting preferably only of these embodiments.

[0031] When an indefinite or definite article is used in reference to a singular noun, e.g., "a", "an", or "the", it includes the plural of that noun unless otherwise specifically stated.

[0032] In the context of the present invention where a numerical value is indicated, those skilled in the art will understand that the technical effect of the feature under discussion is ensured within a precision interval, which typically includes a deviation given as ±10%, preferably ±5% of the numerical value.

[0033] Furthermore, in the specification and claims, the terms first, second, third, (a), (b), (c), etc. are used to distinguish similar elements and not necessarily to describe an order or temporal sequence. It should be understood that the terms so used are interchangeable where appropriate, and the embodiments of the present invention described herein are capable of operating in an order other than that described or illustrated herein.

[0034] Further definitions of the terms will be given where the terms are used hereinafter. The following terms or definitions are provided merely to assist in understanding the present invention. These definitions should not be construed as having a scope less than that understood by a person of ordinary skill in the art.

[0035] In one aspect, the present invention relates to a device for processing a biological sample by means of a centrifugal force applied along a vertical axis of the device, the device comprising:

[0036] (i) at least a first compartment for performing a first step of sample processing, the first compartment comprising a structure for loading a biological sample at its proximal part and a first microfluidic structure at its distal part; and

[0037] (ii) at least a second compartment for performing a second step of sample processing, a proximal end of the second compartment being in fluid communication with a distal end of the first compartment, the second compartment comprising a second microfluidic structure at its distal part, the second step of sample processing being consecutive with the first step of sample processing;

[0038] wherein the first microfluidic structure comprises a capillary having a first average diameter, the second microfluidic structure comprises a capillary having a second average diameter, the first average diameter being greater than the second average diameter, such that a biological sample can be made to pass through the first microfluidic structure by applying a first centrifugal force along the vertical axis of the device, and such that a biological sample can be made to pass through the second microfluidic structure by applying a second centrifugal force along the vertical axis of the device, the second centrifugal force being greater than the first centrifugal force.

[0039] The device of the present invention includes at least two compartments: a first compartment and a second compartment. The device may have more than two compartments, such as three, four, five, six or more compartments. The compartments are arranged vertically such that the distal end of one compartment is connected to the proximal end of the next compartment, so that the two compartments are in fluid communication with each other, i.e., the sample to be processed can be transferred from one compartment to the next compartment. The connection between the two compartments (e.g., at least the first compartment and the second compartment) can be rigid, i.e., the two compartments are equivalent to an integral entity. Alternatively, the connection between the two compartments can be detachable, for example, in the form of a plug connection (i.e., a plug and a socket) or a bolt connection (i.e., a threaded assembly).

[0040] At least the first compartment and the second compartment of the device are equivalent to reaction chambers for performing respective consecutive steps of sample processing. The first processing step is performed in the first compartment, and the second processing step (which is consecutive with the first processing step) is performed in the second compartment, and so on. The compartments can have a spherical, tubular (cylindrical) or conical shape, with or without a taper towards the proximal and / or distal ends. The first compartment includes a structure for loading a biological sample, and the structure is arranged at the proximal end of the compartment. In its simplest form, this structure is an opening through which the biological sample is introduced into the compartment. The opening can be sealed or locked with any form of lid (e.g., a rotary lid). Alternatively, the structure for loading the sample can be configured as a concave surface at the proximal end of the first compartment, which has an opening at the center of the bottom to facilitate the flow of fluid towards the center of the bottom (even for small sample volumes at the μl level), thereby facilitating the application and processing of the complete sample. In some embodiments, the structure for loading is arranged to be close to or directly adjacent to the first microfluidic structure in at least the first compartment.

[0041] Each compartment includes a microfluidic structure in its distal portion, and the microfluidic structure is generally arranged in the distal third of the compartment, particularly within the distal 20% or 10% of the compartment. As used herein, the term "microfluidic structure" refers to a system of one or more microchannels or capillaries in a common polymer matrix. The microfluidic structure can also be directly arranged at the connection site between the two compartments. Generally, the microfluidic structure of the device is configured as a molecular sieve or includes a capillary barrier. The microfluidic structure has a thickness of generally at most 10 mm. In a particular embodiment, the microfluidic structure has a thickness of 0.5 mm, 1 mm, 2 mm, 2.5 mm, 3 mm, 4 mm, 5 mm, 6 mm, 7 mm, 7.5 mm, 8 mm, 9 mm or 10 mm. The flow of fluid through the microfluidic structure is driven by applying an external centrifugal force along the vertical axis of the device. Therefore, in a typical embodiment of the present invention, it is not expected to further involve the arrangement of a micropump for actively manipulating fluid flow. Figure 2Schematically shows the general technical principle on which the device of the present invention is based.

[0042] The microfluidic structure can be produced by various established additive manufacturing techniques (an exemplary review is given), such as 3D printing (Ngo, T.D. et al. (2018) Composites Part B: Engineer. 143, 172 - 196), injection molding (Singh, S. & Verma, A. (2017) Materials today Proc. 4, 1423 - 1433), fused deposition modeling (Penumakala, P.K. et al. (2020) Composites Part B: Engineer. 201, e108336), stereolithography (Huang, J. et al. (2020) Processes 8, e1138), selective laser sintering (Rajesh, R. et al. (2015) Int. J. Curr. Engineer. Sci. Res. 2, 91 - 100) and hot embossing (Deshmukh, S.S. & Goswami, A. (2020) Materials today Proc. 26, 405 - 414). In a specific embodiment, the microfluidic structure is produced by means of 3D printing or by means of injection molding.

[0043] The microfluidic structure includes one or more capillaries that allow fluid to be transferred through the structure. Generally, the one or more capillaries have a tubular (cylindrical) or substantially tubular shape. However, non - cylindrical capillaries are also possible. Generally, the average diameter of the capillaries is in the range between 10 μm and 2 mm, preferably in the range between 50 μm and 1.5 mm or 100 μm and 1 mm. Exemplary ranges of the average diameter are 0.1 mm to 0.5 mm, 0.2 mm to 0.8 mm and 0.5 mm to 1 mm. Exemplary average diameters are 10 μm, 20 μm, 30 μm, 40 μm, 50 μm, 60 μm, 70 μm, 80 μm, 90 μm, 100 μm, 0.2 mm, 0.3 mm, 0.4 mm, 0.5 mm, 0.6 mm, 0.7 mm, 0.8 mm, 0.9 mm, 1 mm, 1.1 mm, 1.2 mm, 1.3 mm, 1.4 mm, 1.5 mm, 1.6 mm, 1.7 mm, 1.8 mm, 1.9 mm and 2 mm.

[0044] The average diameter of the capillaries included in the first microfluidic structure (in the first compartment) is greater than the average diameter of the capillaries included in the second microfluidic structure (in the second compartment). Generally, the average diameter of the capillaries in the first microfluidic structure is 1.5 to 15 times, preferably 2 to 10 times, the average diameter of the capillaries in the second microfluidic structure. In an exemplary embodiment, the average diameter of the capillaries in the first microfluidic structure is 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 times the average diameter of the capillaries in the second microfluidic structure. For example, the average diameter of the capillaries in the first microfluidic structure can be 1 mm and the average diameter of the capillaries in the second microfluidic structure can be 0.1 mm; or, the average diameter of the capillaries in the first microfluidic structure can be 0.5 mm and the average diameter of the capillaries in the second microfluidic structure can be 0.2 mm.

[0045] If the device includes a third (or any other) compartment having a third (or any other) microfluidic structure, the average diameter of the capillaries included in the third (or any other) microfluidic structure is less than the average diameter of the capillaries included in the second (or any previous) microfluidic structure. The above considerations, with necessary modifications, apply.

[0046] The smaller average diameter of the capillaries in the second microfluidic structure compared to the first microfluidic structure results in a higher resistance for the sample to pass through the second microfluidic structure. Therefore, the centrifugal force applied to transfer the sample through the second microfluidic structure must be higher than the centrifugal force applied to transfer the sample through the first microfluidic structure. In other words, if a first centrifugal force is applied along the vertical axis of the device to enable the sample to pass through the first microfluidic structure and a second centrifugal force is applied along the vertical axis of the device to enable the sample to pass through the second microfluidic structure, the second centrifugal force must be greater than the first centrifugal force.

[0047] In a preferred embodiment, the first microfluidic structure exhibits a more hydrophilic property than the second microfluidic structure. To this end, the material used to fabricate the first microfluidic structure can exhibit a more hydrophilic property than the material used to fabricate the second microfluidic structure. Additionally or alternatively, the first microfluidic structure can have a functional surface modification to exhibit a more hydrophilic property than the second microfluidic structure.

[0048] Hydrophilic and hydrophobic materials are usually defined by the geometry of water on a flat surface, particularly by the angle between the edge of a liquid droplet and the surface beneath it. This is commonly referred to as the (static) contact angle. However, more precisely, there is no equilibrium contact angle in nature, and thus no truly static contact angle. Instead, the advancing contact angle (i.e., the highest possible contact angle) and the receding contact angle (i.e., the lowest possible contact angle) should be determined to obtain a true picture of surface wetting and thus the hydrophilic or hydrophobic nature of the material. Generally speaking, if a liquid droplet spreads and wets a large area of the surface, the contact angle is less than 90°, and the surface is considered hydrophilic or water-loving. In contrast, if the liquid droplet forms a sphere that hardly touches the surface, the contact angle is greater than 90°, and the surface is hydrophobic or water-repellent. The higher the polarity of the material or the more charged functional groups it has, the higher its solubility in water or other polar substances (aqueous solvents), and the more hydrophilic it is.

[0049] Generally, the microfluidic structures used herein are made of polymeric materials. Suitable polymeric materials include polypropylene, polyethylene, polytetrafluoroethylene, poly(methyl methacrylate), polycarbonate, polystyrene, polyvinyl chloride, polyimide, polydimethylsiloxane, cyclic block copolymers, cyclic olefin (co)polymers, and combinations thereof. All of these polymeric materials are well-known and commercially available from various suppliers. A person skilled in the art is well aware of the hydrophilic properties of a given polymeric material compared to another polymeric material. For example, polytetrafluoroethylene is significantly more hydrophobic than polypropylene due to its non-polar molecular structure. Thus, a first microfluidic structure made of polypropylene exhibits more hydrophilic properties than a second microfluidic structure made of polytetrafluoroethylene.

[0050] A person skilled in the art is also well aware of methods for surface functionalization (modification) to enhance the hydrophilic properties of a given polymeric material. Suitable methods (an exemplary review is given) include plasma irradiation (e.g., by corona discharge; Agrawal, N.K. et al. (2019) In: Radiation Effects in Polymeric Materials. Kumar, V. et al. (eds.), Springer Press) Chemical vapor deposition (Sun, L. et al. (2021) Nat. Rev. Methods Primer 1, e5), ultraviolet irradiation (Jaganathan, S.K. et al. (2014) J. Materials Sci. 50, 2007 - 2018), and etching (Mijovic, J.S. & Koutsky, J.A. (1977) Polymer Plastics Technol. and Engineer. 9, 139 - 179). For example, plasma irradiation can also be combined with the pre - deposition of hydrophilic polymers such as polyvinyl alcohol or methacrylate compounds).

[0051] Similar to the smaller average diameter of the capillaries in the second microfluidic structure compared to the first microfluidic structure, the second microfluidic structure (in terms of its material) exhibits lower hydrophilic properties, resulting in a higher resistance for the sample to pass through the second microfluidic structure. Therefore, if a first centrifugal force is applied along the vertical axis of the device to enable the sample to pass through the first microfluidic structure and a second centrifugal force is applied along the vertical axis of the device to enable the sample to pass through the second microfluidic structure, the second centrifugal force must be greater than the first centrifugal force.

[0052] In certain embodiments, the device further includes a compartment for sample loading, i.e., a separate entity for introducing a biological sample into the device. Otherwise, at least the first compartment is used for loading the sample. The presence of a separate compartment for sample loading can depend on various factors, such as the volume or viscosity of the sample that may require additional operations before further processing of the sample. However, for many applications, the sample can be directly applied to at least the first compartment. The distal end of the compartment for sample loading can be connected to the proximal end of at least the first compartment such that the two compartments are in fluid communication. The microfluidic structure can be arranged in the distal part of the compartment for sample loading. The connection between the compartment for sample loading and at least the first compartment can be rigid, i.e., the two compartments are equivalent to an integral entity. Alternatively, the connection between the two compartments can be detachable, for example, in the form of a plug connection (i.e., plug and socket) or a bolt connection (i.e., threaded assembly). The compartment for sample loading can have a concave bottom surface to concentrate the sample (along with any buffer or other reagent) at the center of the bottom, thus facilitating complete processing. The opening in the bottom surface for transferring the sample to at least the first compartment can be sealed with a seal having a rupture point that ruptures after the compartment for sample loading is connected to at least the first compartment, for example, by means of a push column arranged at the proximal end of at least the first compartment. The compartment for sample loading has an additional opening for sample loading (e.g., on its top surface). The opening for sample loading can be sealed or locked with a lid in any form (e.g., a rotary lid).

[0053] In other specific embodiments, the device further includes a reagent reservoir, wherein the distal end of the reagent reservoir can be connected to the proximal end of the compartment for sample loading such that the two compartments are in fluid communication. Alternatively, the distal end of the reagent reservoir can be connected to the proximal end of at least the first compartment such that the two compartments are in fluid communication. For many applications, it may be sufficient to directly connect the reagent reservoir to at least the first compartment. The connection between the reagent reservoir and the compartment for sample loading or at least the first compartment can be rigid, i.e., the two compartments are equivalent to a monolithic entity. Alternatively, the connection between the two compartments can be detachable, for example, in the form of a plug connection (i.e., plug and socket) or a bolt connection (i.e., threaded assembly). The reagent reservoir is used to store buffers and / or reagents (e.g., chelating agents, lysis reagents, and enzymes). In a specific embodiment, the reagent reservoir is pre-filled with one or more reagents. The pre-filled reagent reservoir can be sealed, for example, with a lid. The reagent reservoir can have a concave bottom surface to concentrate the reagents (and also the mixture of reagents and biological samples during the operation of the device) at the center of the bottom, thus facilitating complete processing. The opening in the bottom surface for transferring the reagents to the compartment for sample loading or at least the first compartment can be sealed with a seal having a rupture point that ruptures after connection to the compartment for sample loading or at least the first compartment, for example, by means of a push column arranged at the proximal end of the compartment for sample loading or at least the first compartment.

[0054] In still some other specific embodiments, the device further includes a sample collection reservoir for collecting the processed sample, wherein the proximal end of the sample collection reservoir can be connected to the distal end of at least the second compartment (i.e., the terminal compartment for performing the steps of sample processing) such that the two compartments are in fluid communication. The connection between the sample collection reservoir and at least the second compartment can be rigid, i.e., the two compartments are equivalent to a monolithic entity. Alternatively, the connection between the two compartments can be detachable, for example, in the form of a plug connection (i.e., plug and socket) or a bolt connection (i.e., threaded assembly). The sample collection reservoir can have a concave bottom surface to concentrate the (finally) processed sample. The sample collection reservoir can be configured to be connectable to any device for performing any downstream application using the processed sample. Alternatively, in the absence of a separate sample collection reservoir, the terminal compartment of the device (e.g., the second compartment of a device having a first compartment and a second compartment) can have an opening in the bottom surface or no bottom surface, such that the sample can be directly collected in the reaction vessel or other container in which the device is placed.

[0055] The device of the present invention can be used to perform any step of processing a biological sample after obtaining the sample (e.g., obtained directly from a subject or from a storage device or repository) in order to prepare the sample for downstream applications (e.g., nucleic acid amplification and sequencing, mass spectrometry, immunoassay methods, etc.). In a specific embodiment, the processing of the biological sample includes at least partial purification of the sample, i.e., for example, depletion of a part of the materials contained in the sample or removal of any debris.

[0056] In a preferred embodiment, the processing of the biological sample includes at least two steps selected from the group consisting of: sample collection, sample filtration, sample desalting, sample concentration, sample dilution, sample solubilization and / or digestion, depletion of a part contained in the sample, and enrichment of a part contained in the sample. The actual processing steps performed particularly depend on the type of biological sample to be analyzed and the type of downstream application the processed sample is to undergo. The step of collecting the sample can include ensuring the processing of the entire sample volume, for example, by using a structure for sample loading having a concave bottom surface. A person skilled in the art is well aware of selecting appropriate processing steps for a given setup. Such processing steps are widely established in the art and are standard in the field of molecular biology (see, for example, Sambrook, J. et al. (1989) Molecular Cloning: A Laboratory Manual. 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Ausubel, F.M. et al. (2001) Current Protocols in Molecular Biology. Wiley & Sons, Hoboken, NJ).

[0057] Generally, the first processing step involves changing the viscosity of the sample or the concentration of a part of the materials contained in the sample. These steps can also be part of sample collection. Thus, such a first processing step can involve diluting the sample in an appropriate buffer to facilitate subsequent steps (e.g., an undiluted whole blood sample may clog the affinity resin in a subsequent fractionation step). The first processing step can also involve desalting of the biological sample, for example, by using a filter membrane or a semipermeable membrane and reverse osmosis, since a too high salt concentration may interfere with subsequent processing steps. For this purpose, a reaction compartment including a semipermeable membrane can be provided. In another embodiment, the first processing step involves solubilization or lysis of the biological sample, i.e., breaking down tissues and cells to release any (large) molecules of interest, such as nucleic acids or proteins.

[0058] Typically, the second processing step involves fractionation of the materials contained in the biological sample, for example by selective enzymatic digestion of nucleic acids (by addition of nucleases) or proteins (by addition of proteases), or by selective removal (depletion) of any high-abundance macromolecules that may "mask" low-abundance macromolecules and thus obscure the test results. For example, it is well known that just 20 proteins present in a human plasma sample (such as albumin, plasminogen, immunoglobulin G, immunoglobulin M, immunoglobulin A, apolipoproteins) account for 97% to 98% of the total mass of proteins in the plasma. To deplete these proteins, an affinity matrix can be provided to which the proteins selectively bind via specific antibody molecules (or binding fragments thereof). Such affinity matrices for various purposes are commercially available from various suppliers and can be arranged in the reaction compartment of the device.

[0059] Figure 4 An exemplary device according to the invention suitable for processing urine samples is shown. After sample loading, the first processing step is to desalt the sample by filtration through a semi-permeable membrane disposed in the first compartment. After passing through the first microfluidic structure (capillary barrier), the second processing step is to deplete a large amount of proteins by selective capture on an affinity resin or matrix disposed in the second compartment. After passing through the second microfluidic structure (capillary barrier), the processed sample is collected in the sample collection memory.

[0060] In a specific embodiment, the device of the invention is integrally formed as a single unit. At least the first and second compartments for sample processing and, where present, the compartment for sample loading and / or the reagent memory and / or the sample collection memory are equivalent to a single integral entity. In other specific embodiments, at least one compartment of the device is formed as a separate component that can be connected to the remaining one or more compartments of the device. For example, at least the first and second compartments for sample processing can be equivalent to a single integral entity, while the reagent memory and the sample collection memory exist as separate entities. The compartment for sample loading and the reagent memory can also be configured as a single integral entity. It is also possible to provide at least the first and second compartments as separate entities. For example, at least the first and second compartments can be provided with a microfluidic structure having capillaries with a specific predetermined average diameter. Additionally or alternatively, at least the first and second compartments can be provided for a specific step of sample processing, such as a compartment including a specific affinity resin for selectively binding a portion of the biological sample or a compartment including a defined membrane for desalting the biological sample. In other specific embodiments, the device described herein is provided as a kit-of-parts including at least two separate entities, suitable for a specific application where applicable.

[0061] Figure 1 , Figure 4 and Figure 5 An exemplary apparatus according to the invention is shown in FIG. Figure 5 An exemplary workflow for sample processing using this device is also shown.

[0062] In a specific embodiment, the device is configured to be placed in a reaction vessel and used with the reaction vessel, particularly used with a standard reaction tube in a centrifugal process. The volume of a reaction vessel or reaction tube is generally 0.1ml to 50ml, preferably 0.2ml to 10ml, more preferably 0.5ml to 2.5ml. For example, a reaction tube with a volume of 0.5ml, 1.0ml, 1.5ml, 2ml, 5ml, 10ml, 15ml or 50ml can be adopted. This reaction tube can be commercially available from various suppliers. The device can be configured to be placed in a reaction vessel in a manner that is tightly matched or close to a matching module, to reduce or prevent moving in a reaction vessel in a centrifugal process. Alternatively, the device can be configured to directly use similarly to a standard reaction tube, that is, the device is shaped to be similar to this standard reaction tube.

[0063] In another aspect, the invention relates to a method for processing a biological sample by means of a centrifugal force applied along a vertical axis of a processing device, the method comprising:

[0064] (i) introducing a biological sample into a device as defined above;

[0065] (ii) performing a first step of sample processing in at least a first compartment of the device by applying a first centrifugal force along a vertical axis of the device, thereby enabling the sample to pass through a first microfluidic structure; and

[0066] (iii) performing a second step of sample processing in at least a second compartment of the device by applying a second centrifugal force along the vertical axis of the device, thereby enabling the sample to pass through a second microfluidic structure,

[0067] The second centrifugal force is greater than the first centrifugal force.

[0068] A biological sample can be directly introduced into at least a first compartment. Generally, the sample is applied to a structure for sample loading in at least the first compartment. In a particular embodiment where the device includes a compartment for sample loading, the biological sample is introduced into the additional compartment connected to at least the first compartment of the device. The volume of the biological sample generally ranges between 5 μl and 25 ml, depending on the type of sample and the application involved. For example, if plasma or serum is used, a relatively small sample volume in the range of 5 μl to 100 μl or in the range of 10 μl to 50 μl may be sufficient for most applications. If an urine sample is analyzed, the volume may be in the range of 5 ml to 25 ml or in the range of 10 ml to 20 ml. Those skilled in the art are well aware of selecting an appropriate sample volume for a given setting.

[0069] After the biological sample has been introduced into the device, a first step of sample processing can be performed in at least the first compartment. Alternatively, before performing the first step of sample processing, the sample can first be mixed with a suitable buffer and / or other reagents (such as chelating agents, lysis reagents, and enzymes). In a particular embodiment, after the biological sample has been introduced into a device including a reagent reservoir, the reagent is released from the reagent reservoir connected to the compartment for sample loading of the device or connected to at least the first compartment. The reagent can be pre-filled in a sealed reagent reservoir, and the release of the reagent can be achieved by rupturing a seal at the bottom surface of the reagent reservoir at a rupture point, for example, by means of a push column arranged at the proximal end of the compartment for sample loading or at least the first compartment, thereby connecting the reagent reservoir to the device (connecting to the compartment for sample loading of at least the first compartment).

[0070] Then the device (which is optionally in a suitable reaction vessel) is placed in a centrifuge to perform a first step of sample processing in at least the first compartment of the device. For this purpose, a first centrifugal force is applied along the vertical axis of the device, such that the sample can pass through the first microfluidic structure and thus be transferred to at least a second compartment of the device. The transfer can be controlled by the applied centrifugal force. As long as the applied centrifugal force is less than the first centrifugal force required to pass through the first microfluidic structure, the sample will remain in at least the first compartment. Only when the first centrifugal force is reached can the sample pass through the first microfluidic structure.

[0071] The following is entirely within the knowledge of those skilled in the art: calculating the centrifugal force that needs to be applied to cause a liquid to pass through a microfluidic structure of a certain configuration.

[0072] To calculate how much pressure (ΔPa, in N / m 2 2) is required to cause any liquid to pass through a tubular capillary of a given material, the following equation can be used:

[0073]

[0074] where σ is the surface tension in N / m (describing the cohesive force in the liquid), Θ A is the advancing contact angle (see the definition in the legend of Figure 7 ), and D is the diameter of the capillary in m. When additionally considering the angular velocity during the centrifugation process and the diameter of the rotor used for centrifugation (i.e., the distance between the capillary and the rotation center), the required pressure (ΔPd, in N / m 2 ) can be calculated using the following equation:

[0075]

[0076] where ρ is the density of the liquid in kg / m 3 , ω 2 is the angular velocity in rad / s (determining how many 2Pi pass per second), and r is the distance between the capillary and the rotation center in m. Then, the point at which the centrifuge generates the same pressure as that required to drive the liquid through the capillary is determined. This correlation can be expressed by the following equation:

[0077]

[0078] If the equation is solved for ω, the angular velocity required to drive a liquid (with density and surface tension) through a capillary made of any material (with an advancing contact angle) and having any diameter (D) at any distance (r) from the rotation center can be calculated as follows:

[0079]

[0080] Many values of the advancing contact angle, liquid density, and surface tension of various materials can be easily found in the literature or easily determined experimentally. Usually, these values are given at a temperature of 20 °C. The angular velocity is usually given in Rad / s. 1 rad = 2Pi ≈ 6.28. To convert rad / s to revolutions per minute (rpm), the rad / s value must be multiplied by 60 s / 2Pi ≈ 9.55.

[0081] For example, by using the above considerations, the centrifugal force (in rpm) required to drive water through a microfluidic structure made of different polymer materials can be calculated based on the average diameter of the capillaries included in the microfluidic structure. Figure 6 Shows respectively polypropylene ( Figure 6 A) and polytetrafluoroethylene ( Figure 6This relationship for two different polymeric materials of B). The thickness of the microfluidic structure employed in the analysis was 1 mm. Centrifugation was carried out at 20 °C using a 2 ml reaction tube with a rotor of 24 cm in diameter. Water has a surface tension of 72.75 N / m and a density of 999 kg / m 3 . The advancing contact angle between water and polypropylene is 100.7°, and the advancing contact angle between water and polytetrafluoroethylene is 110°.

[0082] From Figure 6 A it can be deduced that 1000 rpm is required to drive water through a capillary with an average diameter of 1 mm, and 2500 rpm is required to drive water through a capillary with an average diameter of 0.1 mm. Thus, when applied to the device according to the present invention, the transfer of the sample from one compartment to the next can be precisely controlled by the configuration of the microfluidic structures included in each compartment. Using the above example, if the first microfluidic structure with an average diameter of 1 mm capillary can be traversed by the centrifugal force generated at 1000 rpm, and the second microfluidic structure with an average diameter of 0.1 mm capillary can be traversed by the centrifugal force generated at 2500 rpm, then the fluid flow can be controlled only by changing the average diameter of the capillary. If the material of the first microfluidic structure is further modified by surface functionalization to increase the hydrophilic property, the difference in the corresponding centrifugal forces required to drive water through the first and second microfluidic structures will become more significant.

[0083] From Figure 6 B it can be deduced the further influence of the polymeric material used to fabricate the microfluidic structure. When using polytetrafluoroethylene which is significantly more hydrophobic than polypropylene, the centrifugal force generated at 3500 rpm (only 2500 rpm was required when using polypropylene) can be used to drive water through a capillary with an average diameter of 0.1 mm.

[0084] Furthermore, Figure 7 shows the influence of the advancing contact angle on the following centrifugal forces (in rpm): the centrifugal forces required to transfer water through capillaries with an average diameter of 100 μm (upper curve) and 1 mm (lower curve) (made of an arbitrary polymer such as polytetrafluoroethylene). Again, the thickness of the microfluidic structure employed in the analysis was 1 mm. Centrifugation was carried out at 20 °C using a 2 ml reaction tube with a rotor of 24 cm in diameter. It is evident from the results that as the hydrophobicity increases, the centrifugal force required to drive water through capillaries with a decreasing average diameter increases more than proportionally.

[0085] Returning to the method of the present invention, after the sample has passed through the first microfluidic structure and entered at least the second compartment, the second step of sample processing is performed in at least the second compartment of the device by applying a second centrifugal force along the vertical axis of the device, such that the sample can pass through the second microfluidic structure, wherein the second centrifugal force is greater than the first centrifugal force. The transfer can be controlled by the applied centrifugal force. As long as the centrifugal force is less than the second centrifugal force, the sample will remain in at least the second compartment. Only when the second centrifugal force is reached can the sample pass through the second microfluidic structure. Due to the head-to-tail arrangement of the various compartments of the device, the sample is transferred unidirectionally (i.e., always in the same direction) along the vertical axis of the device. Therefore, the centrifugal force is also applied unidirectionally along the vertical axis of the device.

[0086] If required by a particular protocol, the third step of sample processing can be performed in a third compartment that is in fluid communication with the second compartment. The third compartment is connected to the distal end of the second compartment at its proximal end. A third centrifugal force is applied along the vertical axis of the device, such that the sample can pass through the third microfluidic structure included in the third compartment, wherein the third centrifugal force is greater than the second centrifugal force.

[0087] In a particular embodiment of the present invention, a device is provided that includes compartments (for performing steps of sample processing) having microfluidic structures arranged therein in a predetermined configuration. For example, respectively, the first compartment includes a first microfluidic structure that can be passed through by a first centrifugal force generated at 50 rpm, the second compartment includes a second microfluidic structure that can be passed through by a second centrifugal force generated at 500 rpm, and the third compartment includes a third microfluidic structure that can be passed through by a third centrifugal force generated at 5000 rpm.

[0088] After the processing of the sample has been completed, the processed sample is removed from the device and subjected to downstream applications. To remove the processed sample, at least the second compartment can have an opening at the bottom third of the compartment or directly at the bottom surface. The processed sample can be collected in a reaction tube in which the device is placed for centrifugation. In a particular embodiment, the processed sample is collected in a sample collection memory connected to at least the second compartment of the device.

[0089] The biological sample to be analyzed in this article is a sample derived from a mammal, such as a mouse, rat, hamster, rabbit, cat, dog, pig, cow, horse, or monkey, preferably a human. Such samples can include body tissues (e.g., biopsies or excisions, tissue samples, tissue lysates), swabs (e.g., nasopharyngeal or oropharyngeal swabs, wound swabs, skin swabs), fecal samples, and body fluids (including liquid biopsies), such as blood (whole blood, plasma, serum), saliva, sputum, sweat, urine, cerebrospinal fluid, interstitial fluid, tears, lymph fluid, and aqueous humor. The sample can contain single cells, cell populations (i.e., two or more cells), or cell extracts or suspensions derived from body tissues or swabs, and can be used in an unpurified form or undergo any enrichment or purification steps before use. Those skilled in the art are well aware of various such purification methods (see, e.g., Sambrook, J. et al. (1989) Molecular Cloning: A Laboratory Manual. 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Ausubel, F.M. et al. (2001) Current Protocols in Molecular Biology. Wiley & Sons, Hoboken, NJ). The term "whole blood" refers to blood with all its components (i.e., blood cells and plasma). The term "plasma" denotes the liquid medium of blood. The term "serum" refers to plasma from which the clotting proteins have been removed. In certain embodiments, the biological sample is selected from the group consisting of blood, plasma, serum, saliva, urine, nasopharyngeal swabs, oropharyngeal swabs, and tissue samples.

[0090] In another aspect, the present invention relates to the use of the device as defined above for processing biological samples to be subjected to so-called "omics" applications. "Omics" applications are relatively new biomarker discovery and monitoring tools that can be applied to study large numbers of biomolecules. Generally speaking, the purpose of "omics" applications is to identify, characterize, and quantify all biomolecules of a specific type involved in the structure, function, and dynamics of cells, tissues, or organisms. Many "omics" applications are well known in the art, such as genomics, epigenomics, phenomics, transcriptomics, proteomics, lipidomics, glycomics, microbiomics, and metabolomics.

[0091] Genomics studies the structure, function, evolution, mapping, and editing of genomes with the aim of characterizing and quantifying genes. Epigenomics studies the supporting structures of genomes, including protein and RNA binders, alternative DNA structures, and DNA modifiers. Phenomics is the systematic study of the traits that make up a phenotype. Transcriptomics analyzes the transcriptome of an organism (the sum of all its mRNA transcripts). In addition to molecular identification, it also includes the amount or concentration of each RNA molecule. The term proteome refers to the sum of all proteins in a cell, tissue, or organism. Proteomics is the science of studying the related biochemical properties and functional roles of these proteins and how their amounts, modifications, and structures change during growth and in response to internal and external stimuli. Lipidomics studies all the lipids of an organism, and glycomics studies all the sugars and carbohydrates. The microbiome is the microbial community that can typically coexist in a specific growth environment. The metabolome represents the collection of all metabolites (which are the end products of cellular processes) in a biological cell, tissue, organ, or organism. Metabolomics is the science of studying all the chemical processes involving metabolites.

[0092] In certain embodiments, the device is for processing a biological sample to be subjected to an "omics" application selected from the group consisting of genomics, transcriptomics, proteomics, lipidomics, glycomics, microbiomics, and metabolomics.

[0093] The present invention illustratively described herein can be suitably implemented in the absence of any one or more elements or one or more limitations not specifically disclosed herein. Thus, for example, the terms "comprising," "including," "containing," etc. should be construed broadly and without limitation. Additionally, the terms and expressions used herein have been used in an illustrative rather than a restrictive sense, and in using these terms and expressions, there is no intention to exclude any equivalents of the features shown and described or portions thereof, and it should be recognized that various modifications can be made within the scope of the claimed invention. Accordingly, it should be understood that although the present invention has been specifically disclosed by way of embodiments and optional features, those skilled in the art can adopt modifications and variations of the invention embodied therein, and such modifications and variations are considered to be within the scope of the present invention.

[0094] The present invention has been described herein in a broad and general manner. Each of the narrower species and general sub - groups falling within the general disclosure also forms part of the present invention. This includes the general description of the present invention, as well as conditional or negative limitations removing any subject matter from the genus, whether or not the excised material is specifically recited herein.

[0095] Other embodiments are within the scope of the appended claims. In addition, where features or aspects of the invention are described in terms of a Markush group, those skilled in the art will recognize that the invention is thereby also described in terms of any single member or subgroup of members of the Markush group.

Claims

1. A device for processing a biological sample, which processes the biological sample by means of a centrifugal force applied along a vertical axis of the device, the device comprising: (i) at least a first compartment for performing a first step of sample processing, the first compartment comprising a structure for loading a biological sample at its proximal part and a first microfluidic structure at its distal part; and (ii) at least a second compartment for performing a second step of sample processing, a proximal end of the second compartment being in fluid communication with a distal end of the first compartment, the second compartment comprising a second microfluidic structure at its distal part, the second step of sample processing being consecutive to the first step of sample processing; wherein the first microfluidic structure comprises a capillary having a first average diameter, the second microfluidic structure comprises a capillary having a second average diameter, the first average diameter being greater than the second average diameter, such that a biological sample can be made to pass through the first microfluidic structure by applying a first centrifugal force along the vertical axis of the device, and such that a biological sample can be made to pass through the second microfluidic structure by applying a second centrifugal force along the vertical axis of the device, the second centrifugal force being greater than the first centrifugal force.

2. The device according to claim 1, wherein The first microfluidic structure exhibits a more hydrophilic property than the second microfluidic structure.

3. The device according to claim 2, wherein The material for manufacturing the first microfluidic structure exhibits a more hydrophilic property than the material for manufacturing the second microfluidic structure; and / or wherein the first microfluidic structure has a functional surface modification to exhibit a more hydrophilic property than the second microfluidic structure.

4. The device according to any one of claims 1 to 3, wherein, The processing of the biological sample includes at least partial purification of the sample.

5. The device according to any one of claims 1 to 4, wherein The processing of the biological sample includes at least two steps selected from the group consisting of: sample collection, sample filtration, sample desalting, sample concentration, sample dilution, sample solubilization and / or digestion, depletion of a part contained in the sample, and enrichment of a part contained in the sample.

6. The device according to any one of claims 1 to 5, wherein The device is integrally formed as a single unit; or wherein at least one compartment of the device is formed as a separate component that can be connected to the remaining one or more compartments of the device.

7. The device according to any one of claims 1 to 5, wherein The device is configured to be placed in a reaction vessel and used together with the reaction vessel, the reaction vessel having a volume of 0.1 ml to 50 ml, preferably 0.2 ml to 10 ml, more preferably 0.5 ml to 2.5 ml.

8. The device according to any one of claims 1 to 7, further comprising a compartment for sample loading, a distal end of the compartment for sample loading being connectable to a proximal end of the at least first compartment such that the two compartments are in fluid communication.

9. The device according to any one of claims 1 to 8, further comprising a reagent reservoir, a distal end of the reagent reservoir being connectable to a proximal end of the compartment for sample loading such that the two compartments are in fluid communication; or a distal end of the reagent reservoir being connectable to a proximal end of the at least first compartment such that the two compartments are in fluid communication, in particular wherein the reagent reservoir is pre-filled with a reagent.

10. The device according to any one of claims 1 to 8, further comprising a sample collection memory for collecting the processed sample, the proximal end of the sample collection memory being connectable to the distal end of the at least second compartment such that the two compartments are in fluid communication.

11. A method for processing a biological sample, which processes a biological sample by means of a centrifugal force applied along the vertical axis of the processing device, the method comprising: (i) introducing the biological sample into the device as defined in any one of claims 1 to 10; (ii) performing a first step of sample processing in at least a first compartment of the device by applying a first centrifugal force along the vertical axis of the device, such that the sample can pass through the first microfluidic structure; and (iii) performing a second step of sample processing in at least a second compartment of the device by applying a second centrifugal force along the vertical axis of the device, such that the sample can pass through the second microfluidic structure, wherein the second centrifugal force is greater than the first centrifugal force.

12. The method according to claim 11, further comprising: loading the biological sample into a compartment for sample loading connected to at least a first compartment of the device; and / or releasing a reagent from a reagent memory connected to the compartment for sample loading of the device or connected to at least the first compartment after the biological sample has been introduced into the device.

13. The method according to claim 11 or 12, further comprising: collecting the processed sample in a sample collection memory connected to at least a second compartment of the device.

14. The method according to any one of claims 11 to 13, wherein The biological sample is selected from the group consisting of blood, plasma, serum, saliva, urine, nasopharyngeal swab, oropharyngeal swab, and tissue sample.

15. Use of the device as defined in any one of claims 1 to 10 for processing a biological sample to be subjected to an "omics" application, in particular wherein the "omics" application is selected from the group consisting of genomics, transcriptomics, proteomics, lipidomics, glycomics, microbiomics, and metabolomics.

Images