Connector, marker, capture construct and data storage device

Abstract

There is provided a linker (100, 102) for generating a biological structure (400, 500, 600, 700), comprising: a protein scaffold (106); a first reactive interactor (108) arranged towards a first end of the protein backbone (106) and configured to covalently bind to the second reactive interactor (116); and a first affinity interactor (110) disposed towards the second end of the protein backbone (106) and configured to bind to the second affinity interactor (114). The protein backbone (106) includes a cleavage site (104) between a first end and a second end of the protein backbone (106). Furthermore, one of the first reactive interactor (108) and the first affinity interactor (110) is configured to bind to a first affinity reagent (402) comprising a corresponding second reactive interactor (116) or second affinity interactor (114), and the other of the first reactive interactor (108) and the first affinity interactor (110) is configured to bind to a marker (300) or a second affinity reagent (702) comprising a corresponding second reactive interactor (116) or second affinity interactor (114). In another aspect, markers, data storage devices, and capture constructs including the linker are provided, as well as methods and devices for generating these.

Description

Technical Field

[0001] The present invention relates to a connector for a marker, a capture construct or a data storage device, and methods and devices to generate a marker, a capture construct or a data storage device. Background Art

[0002] Generating structures from biological components or building blocks has received increasing attention. In particular, the high specificity and high affinity of biological components such as antibodies or oligonucleotides are properties that can be exploited when building biological structures from these components (such as markers).

[0003] To give a specific example: space biology is an emerging field of microscopy in which different strategies are used to image biological samples (such as tissue sections) with high spatial resolution and analyze a large number of markers simultaneously. In order to mark a large number of targets in the sample, the number of markers required in this application can be anywhere from about 100 to 10,000. This requires cyclic staining, imaging, and a blanking process, as well as the construction of a large library of markers before their use. Compared to most current microscopy tests (such as immunofluorescence or FISH experiments, which involve only a small number of markers), these new methods require significantly more time to stain the sample, image the sample, and process and analyze the data. In addition, they require significantly more expensive reagents, such as markers.

[0004] A particular problem remains the rapid and efficient generation of large libraries of biological structures, such as markers, with various properties. Summary of the invention

[0005] Therefore, an object of the present application is to provide components for efficiently generating biological structures.

[0006] The above objects are achieved by the subject matter of the independent claims. Advantageous embodiments are defined in the dependent claims and in the following description.

[0007] On the one hand, a connector is provided for generating, particularly assembling a biological structure, the connector comprising: a protein backbone; a first reactive interactor, which is arranged toward the first end of the protein backbone or at the first end of the protein backbone, and is configured to covalently and specifically bind to a second reactive interactor; and a first affinity interactor, which is arranged toward the second end of the protein backbone or at the second end of the protein backbone, and is configured to non-covalently and specifically bind to a second affinity interactor. The protein backbone includes a cleavage site between the first end and the second end of the protein backbone. In addition, one of the first reactive interactor and the first affinity interactor is configured to bind to a first affinity reagent including a corresponding second reactive interactor or a second affinity interactor, and the other of the first reactive interactor and the first affinity interactor is configured to bind to a label or a second affinity reagent including a corresponding second reactive interactor or a second affinity interactor.

[0008] The linker allows two molecules and / or biological building blocks to be selectively linked to each other in an efficient manner, which preferably results in a linkage with little spontaneous dissociation, but is compatible with the introduction of a cleavage site that allows the user to cleave the linkage at their discretion. The linker with two pairs of interactors can be particularly useful for enabling rapid, cost-effective production of large libraries of markers, wherein the marker is a conjugate of a label and an affinity agent, and wherein the label is a construct carrying one or more labeling agents, such as a fluorescent dye or a combination of fluorescent dyes.

[0009] Preferably, the backbone is specifically cleavable by an enzyme (particularly a protease), by cleavage light or by a change in temperature. This enables flexible separation of the ends of the linker, for example when the linker is used in a method with iterative steps, which requires repeated removal of parts of the linker.

[0010] Preferably, the first reactive interactor and the second reactive interactor are configured to form a bioconjugate. For example, the reactive interactor pair may be a SpyTag / SpyCatcher. This enables the reactive interactor pair to have high specificity and thus enables the reactive interactors to have particularly robust covalent binding.

[0011] Preferably, the first affinity interactor is one of streptavidin and biotin, and the second affinity interactor is the other of streptavidin and biotin. This enables the affinity interactor pair to have high affinity and high specificity, and thus enables the affinity interactors to bind robustly.

[0012] Preferably, the streptavidin is a tetramer with one active biotin binding site or a monomer with one active biotin binding site. This allows the affinity interactor pair to bind in a predictable stoichiometric ratio.

[0013] Preferably, the linker is a fusion protein comprising the first affinity interactor, the protein backbone, the cleavage site and the second affinity interactor. This enables particularly efficient production of the linker.

[0014] Preferably, the first affinity interactor is streptavidin and the first affinity reagent comprises an oligonucleotide of the biotin. In particular, the first affinity reagent is a biotinylated oligonucleotide. This enables particularly easy production of the first affinity reagent.

[0015] Preferably, the linker comprises the first affinity reagent. In particular, the first affinity reagent is the biotinylated oligonucleotide, which is configured to bind to the streptavidin of the protein backbone with high affinity.

[0016] On the other hand, a marker for analyzing a biological sample is provided, the marker comprising: the marker bound to the above-described linker, particularly comprising at least one dye. For example, the marker can be attached to a target analyte in the biological sample via the first affinity reagent. It is particularly effective to generate the marker from individual components (such as the linker, the marker, and the first affinity reagent). In addition, a variety of markers can be generated from these individual components with different properties. For example, markers with different optical properties can be used to effectively generate markers with these different optical properties.

[0017] Preferably, the label comprises the second reactive interactor covalently bound to the first reactive interactor of the linker, in particular to form a bioconjugate. This enables a particularly robust attachment of the label to the linker.

[0018] Preferably, the marker comprises a third affinity reagent, the third affinity reagent comprises an oligonucleotide, the oligonucleotide is at least partially complementary to the first affinity reagent, in particular a biotinylated oligonucleotide, of the linker, and hybridizes with the first affinity reagent. Thus, the biotinylated oligonucleotide or the third affinity reagent can bind to a target molecule in a biological sample.

[0019] Preferably, the third affinity reagent further comprises an antibody. The antibody can bind to a target molecule in a biological sample. This enables binding to a variety of target molecules with particularly high specificity.

[0020] Preferably, the marker comprises: an oligonucleotide base comprising the second reactive interactant; and at least one oligonucleotide-coupled dye hybridizing to the oligonucleotide base via complementary base pairing. This enables particularly efficient generation of the marker.

[0021] On the other hand, a capture construct for capturing an analyte of a biological sample is provided, the capture construct comprising: a DNA origami backbone having at least one capture region; at least one affinity capture reagent, the at least one affinity capture reagent comprising the connector described above and the second affinity reagent. The first affinity reagent of the connector binds the affinity capture reagent to the capture region of the DNA origami backbone, and the first affinity reagent is a staple chain of the DNA origami backbone, preferably including the second affinity interactor. In addition, the second affinity reagent is configured to capture an analyte of the biological sample, and preferably, the second affinity reagent includes the second reactive interactor and is covalently bound to the first reactive interactor of the connector. The capture construct is capable of capturing a plurality of analytes at a predetermined position on the backbone, the capture region. In addition, the capture construct is capable of capturing the analytes at a particularly high density. The capture of the plurality of analytes enables the analysis of the plurality of analytes.

[0022] Preferably, the DNA origami backbone comprises at least a first orientation indicator and a second orientation indicator. The first orientation indicator and the second orientation indicator can be, for example, fluorescent dyes attached to the backbone. These orientation indicators can determine the spatial orientation or directionality of the capture construct, in particular, the backbone. In this way, the orientation indicators can achieve spatial encoding. This means that different positions on the backbone can be assigned to capture areas reactive to different analytes.

[0023] Preferably, the DNA origami backbone comprises nucleic acids. The size of these DNA origami structures can range from a few nanometers to the micrometer range. To make such DNA origami-based structures, longer DNA molecules (scaffold chains) are folded by so-called staple chains at precisely identified positions. The DNA origami can be designed to provide a self-assembly backbone of a specific predetermined shape. This enables easy and reproducible synthesis and assembly of the backbone. The staple chains can be functionalized position-selectively. In this case, the position resolution is limited to the size of the nucleotides in the nanometer or sub-nanometer range. This has been used in the prior art to generate fluorescent standards in which fluorescent dyes are attached to precisely positioned bands on the DNA origami. These standards are called "nanorulers" and are used to calibrate imaging systems such as confocal or super-resolution microscopes (e.g. STED), for example, as disclosed in US2014 / 0057805 A1.

[0024] The DNA origami framework provides a scaffold for the affinity capture reagent. Preferably, the DNA origami structure comprises at least one scaffold chain and a plurality of staple chains, wherein the staple chains are complementary to at least part of the scaffold chains, and the staple chains are configured to cause the scaffold chains to form a predetermined conformation. In particular, these chains are oligonucleotides. This enables self-assembly to generate a skeleton having a predetermined two-dimensional or three-dimensional shape. In addition, this enables the capture region to be placed at a specific site on the skeleton. Preferably, the affinity capture reagent of the capture region can be attached to the staple chains of the skeleton at a predetermined position. The staple chains allow precise functionalization of the DNA origami space at their corresponding positions on the DNA origami. Therefore, each capture region can be located at a specific staple chain along the skeleton or a group of staple chains close to each other. Since the staple chains are located at predetermined positions, the position of the capture region can also be predetermined.

[0025] Further details of the capture construct are disclosed in application PCT / EP2022 / 058640. The contents of this application are incorporated herein by reference in their entirety.

[0026] In another aspect, a data storage device is provided, comprising: a DNA origami backbone having a plurality of attachment sites; the label bound to the connector, the label particularly comprising at least one dye. The first affinity reagent is an oligonucleotide staple strand of the DNA origami backbone, which preferably comprises the second affinity interactor, and the first affinity reagent comprises a unique oligonucleotide sequence, the unique oligonucleotide sequence being configured to bind the connector having the label to a complementary sequence of one of the attachment sites.

[0027] Preferably, the DNA origami backbone of the data storage device comprises at least a first orientation indicator and a second orientation indicator. The first orientation indicator and the second orientation indicator can be, for example, fluorescent dyes attached to the backbone. These orientation indicators can determine the spatial orientation or directionality of the capture construct (in particular, the backbone). In this way, the orientation indicator achieves spatial encoding. This means that different positions on the backbone can be assigned to capture areas that are reactive to different analytes.

[0028] The attachment site is a unique nucleic acid sequence, preferably a unique nucleic acid sequence of the staple chain. Preferably, the marker can be attached to the staple chain of the skeleton at a predetermined attachment site. Since the staple chain is located at a predetermined position, the position of the attachment site can also be predetermined.

[0029] By providing or generating the DNA origami backbone in a specific or predetermined manner, the position of the attachment sites for the markers relative to the backbone and / or at least one first orientation indicator and one second orientation indicator is predetermined or known. Thus, by providing a suitable or predetermined plurality of markers to attach to corresponding attachment sites, information can be stored in the data storage device.

[0030] Further details of the data storage device are disclosed in application EP22191690.1. The content of this application is incorporated herein by reference in its entirety.

[0031] In yet another aspect, a method for generating the marker is provided, comprising the steps of: binding the first affinity reagent to the linker, in particular, binding the biotinylated oligonucleotide to the linker; and binding the linker to the label, in particular, forming a bioconjugate.

[0032] Preferably, the method further comprises the following steps: combining the first affinity reagent with the third affinity reagent. In particular, the first affinity reagent is the biotinylated oligonucleotide, and the third affinity reagent is an oligonucleotide coupled to an antibody having a sequence partially complementary to the biotinylated oligonucleotide. In this step, the biotinylated oligonucleotide is bound to the oligonucleotide of the antibody.

[0033] In another aspect, a method for generating the capture construct or for generating the data storage device is provided, the method comprising the steps of combining the first affinity reagent with the connector, combining the connector with the label or the second affinity reagent, and combining the first affinity reagent with the DNA origami backbone. This enables particularly efficient generation of the capture construct or the storage device.

[0034] In another aspect, a device for generating a marker, a capture construct or a data storage device is provided, the device comprising: a liquid dispensing unit; a plurality of liquid containers; and a control unit, the control unit being configured to instruct the liquid dispensing unit to dispense a liquid containing a marker, a first affinity reagent, a second affinity reagent, a third affinity reagent or a DNA origami skeleton into the liquid container according to the method steps described above. This enables particularly efficient generation of the marker, the capture construct or the data storage device.

[0035] Preferably, the liquid dispensing unit is configured to dispense the liquid using acoustic droplet ejection technology. This enables particularly precise dispensing of small volumes. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Specific embodiments will be described below with reference to the accompanying drawings, in which: Figure 1 is a schematic diagram of the linker in the states before and after cleavage of the cleavage site; Figure 2 is a schematic diagram of several configurations of streptavidin; Figure 3 is based on Figure 2 A schematic diagram of a nucleic acid backbone having sequencing information of a target nucleic acid strand attached thereto; Figure 4 is a schematic diagram of a label including a second reactive interactor; Figure 5 is a schematic diagram of the steps used to assemble the marker; Figure 6 is a schematic diagram of steps for assembling a data storage device; and Figure 7 is a schematic diagram of the steps used to assemble the capture construct. DETAILED DESCRIPTION

[0037] Figure 1 1 is a schematic diagram of a connector 100, 102 before and after cleavage of a cleavage site 104 of the connector 100, 102. The connector 100, 102 comprises a protein backbone 106, which comprises a cleavage site 104. A connector first reactive interactant 108 is disposed at a first end of the connector 100, 102, and a connector first affinity interactant 110 is disposed at a second end of the connector 100, 102. Preferably, both the first affinity interactant 110 and the first reactive interactant 108 are proteins. Therefore, the protein backbone 106 and the interactants 110, 108 can be generated by expression of a single expression cassette.

[0038] For example, the cleavage site 104 can be a specific motif that can be cleaved by the protease 112. Alternatively, the cleavage site 104 can be light or temperature sensitive, so that the protein backbone 106 is cleaved by light or temperature changes. After the cleavage site 104 is cleaved, the first end of the corresponding connector 100, 102 and the second end of the corresponding connector 100, 102 are separated from each other. This cleavage can be irreversible.

[0039] The first reactive interactant 108 is configured to irreversibly and covalently bind to the second reactive interactant 116. Examples of these pairs of reactive interactants 108, 116 include proteins that form bioconjugates, such as Tag / Catcher pairs, for example SpyTag / SpyCatcher. Thus, for example, as in the connector 100, the first reactive interactant 108 can be a Tag portion, and the second reactive interactant 116 can be a Catcher portion. However, alternatively, as in the connector 102, the first reactive interactant 108 can be a Catcher portion, and the second reactive interactant 116 can be a Tag portion.

[0040] The first affinity interactor 110 is configured to specifically bind to the second affinity interactor 114. The first affinity interactor 110 and the second affinity interactor 114 specifically bind to each other based on intermolecular forces (such as ionic bonds, hydrogen bonds and van der Waals forces), and the binding is reversible. A particularly preferred example of these paired affinity interactors 110, 114 is biotin and streptavidin that bind to each other with very high affinity. The binding of biotin to streptavidin is irreversible under physiological conditions. Similar to the first reactive interactor 108 and the second reactive interactor 116, the first affinity interactor 110 can be one of biotin and streptavidin, and the second affinity interactor 114 can be another of biotin and streptavidin.

[0041] Thus, linkers 100 , 102 are capable of specific binding to two further entities, either comprising one of affinity interactants 110 , 114 or one of reactive interactants 108 , 116 , while also enabling subsequent separation via cleavage site 104 .

[0042] Figure 2 Schematic diagrams of several configurations 200, 202, 204, 206, 208 of streptavidin. The first configuration 200 of streptavidin is a natural tetramer with four active biotin binding sites 210. The second configuration 202 of streptavidin is a tetramer with three active sites 210 and one non-functional biotin binding site 212. The third configuration 204 of streptavidin is a tetramer with two active sites 210 and two non-functional biotin binding sites 212. The fourth configuration 206 of streptavidin is a tetramer with one active site 210 and three non-functional biotin binding sites 212. The fifth configuration 208 of streptavidin is a monomer with one active site 210.

[0043] By varying the number of active sites 210, one can vary the number of entities that can bind to the linkers 100, 102. Furthermore, by selecting a configuration 206, 208 with only one binding site, the stoichiometric ratio of the linkers 100, 102 and the entities that bind to each other via the first affinity interactor 110 and the second affinity interactor 114 is predictable, such that only one linker 100, 102 binds to one entity.

[0044] Figure 3is a schematic diagram of a label 300 including a second reactive interactant 116. This enables the label 300 to be bound to the linker 100. Alternatively, the label 300 may include any of the other first reactive interactants 108, first affinity interactants 110, or second affinity interactants 114, so as to bind the label 300 to a linker including a corresponding pair of matching interactants 108, 110, 114, 116.

[0045] The marker 300 includes a dye. Figure 3 The shape marked with reference numeral 302 in the figure represents the dye. The marker 300 includes five individual fluorescent dyes 304, 306, 308, 310, 312. These dyes differ in their optical properties (such as excitation wavelength, emission wavelength and emission lifetime). The dyes 304, 306, 308, 310, 312 are each coupled to an oligonucleotide that is configured to hybridize with a specific sequence on a central nucleic acid backbone 314. The central nucleic acid backbone 314 is in turn configured to hybridize with a specific oligonucleotide 316 that includes a second reactive interactant 116.

[0046] The marker 300 can be assembled flexibly from individual parts 304, 306, 308, 310, 312, 314, 316. For example, in order to assemble multiple unique markers, multiple individual dyes 304, 306, 308, 310, 312 coupled to oligonucleotides can be combined with a central nucleic acid backbone 314 and an oligonucleotide 316 comprising a second reactive interactor 116. Unique base pairing between the individual oligonucleotides enables the assembly of a predetermined marker. For example, the markers can differ in the optical properties of the individual dyes attached to the central nucleic acid backbone 314.

[0047] Figure 4 is a schematic diagram of the steps for assembling a marker 400. The marker 400 can be generated by combining the marker 300 with the linker 100 and the first affinity reagent 402. The marker 300 including the second reactive interactant 116 is bound to the first reactive interactant 108 of the linker 100 to form a bioconjugate. The first affinity reagent 402 including the second affinity interactant 114 is bound to the first affinity interactant 110 of the linker 100. The first affinity reagent 402 can be, for example, an oligonucleotide 404 having a biotin 114 portion. Due to the specificity of each interactant 108, 110, 114, 116 to the corresponding counter interactant 108, 110, 114, 116, the marker 400 can be assembled in an efficient manner.

[0048] Based on the specific affinity of the first affinity reagent 402, the marker 400 can be introduced into the biological sample to mark a specific biological structure. For example, the first affinity reagent 404 of the marker 400 can mark a genetic structure 406 that binds to it via base pairing. Therefore, when the fluorescence of the dye 302 in the sample is detected, for example by microscopy, the location of the dye 302 reveals the location of the structure 406 in the sample.

[0049] When assembling the markers 400, a mix-and-match approach can be employed. Figure 3 As described, a specific marker having a specific optical property can be effectively linked to a specific first affinity reagent 402 having an affinity for a specific biological structure in the biological sample. In this way, a specific marker 400 can be generated simply by combining a solution with a specific marker 300, a linker 100, and a specific first affinity reagent 402 in a container. At the same time, different markers can be generated by combining the solution with another marker, a linker, and another first affinity reagent (particularly, another first affinity reagent having an affinity for another biological structure in the biological sample) in a container. Therefore, several different markers having various optical properties and having an affinity for a specific biological structure in the biological sample can be effectively generated.

[0050] Figure 5 5 is a schematic diagram of a marker 500 having another affinity reagent 502. The other affinity reagent 502 can be an oligonucleotide 504 coupled to an antibody. The marker 500 can be assembled by combining the marker 400 with the other affinity reagent 502 and the oligonucleotide 504 configured to hybridize with the first affinity reagent 404. The marker 500 can then be used to label a biological structure in a biological sample that has an affinity for the other affinity reagent 502. Figure 4 As explained, by mixing and matching marker 400 with a specific label having another affinity reagent 502, multiple markers with specific affinities and optical properties can be efficiently assembled.

[0051] Figure 6is a schematic diagram of the steps for assembling a data storage device 600. The data storage device 600 is assembled from a marker 400 and a DNA origami backbone 602. The DNA origami backbone 602 has a plurality of attachment sites 604. The marker 400, in particular the first affinity reagent 404 of the marker 400, can hybridize with one of the attachment sites 604 of the backbone 602. Each attachment site 604 of the backbone 602 has a unique oligonucleotide sequence, and the first affinity reagent 404 of each marker 400 includes a complementary oligonucleotide sequence. Thus, a plurality of markers, each having a unique complementary sequence, can be attached to a particular backbone 602. By placing the marker at a specific attachment site and by using markers with different optical properties, information can be stored. The connector 100 is capable of efficiently assembling a data storage device 600 by assembling a variety of different markers 400 and combining them with the DNA origami backbone 602.

[0052] Figure 7 is a schematic diagram of steps for assembling a capture construct 700. The capture construct includes a second affinity reagent 702, a connector 100, a first affinity reagent 402, and a DNA origami backbone 704. The capture construct 700 is assembled by combining the second affinity reagent 702, the connector 100, and the first affinity reagent 402 to assemble an affinity capture reagent 706. As explained above, the corresponding interactors 108, 110, 114, 116 are able to effectively assemble the various parts. The capture construct 700 includes at least one capture region 708 that is bound to the affinity capture reagent 706 (particularly, the first affinity reagent 404). The capture region 708 can include multiple affinity capture reagents 706. The second affinity reagent 702 can be a nanobody, such as a nanobody with affinity for a specific target analyte of a biological sample, and includes a second reactive interactor 116. Thus, when the capture construct 700 is added to the biological sample, the target analyte is captured by the capture construct 700, for example, so that the target analyte can be further analyzed. The connector 100 can effectively assemble the capture construct 700 by effectively assembling various different affinity capture reagents 706 and binding them to the respective capture regions 708 of the DNA origami backbone 704. The capture regions 708 can be different from each other because the corresponding affinity capture reagents 706 have a second affinity reagent 702 with affinity for a specific target analyte.

[0053] To assemble the markers 400, 500, data storage devices 600, or capture constructs 700, each of their respective individual components can be provided in a liquid solution, and by combining the solutions, such as by a liquid dispensing unit, the respective interactors 108, 110, 114, 116 of the linkers 100, 102 are brought together. By mixing and matching different first and second affinity reagents, that is, by combining solutions of different first and second affinity reagents, multiple markers 400, 500, data storage devices 600, or capture constructs 700 can be efficiently assembled from these individual components.

[0054] In all drawings, elements having the same or similar functions are indicated by the same reference numerals. As used herein, the term "and / or" includes any and all combinations of one or more of the related listed items and may be abbreviated as " / ". The various features of the embodiments and all combinations of the various features of the embodiments with each other and with the individual features or feature groups of the preceding description and / or claims are considered to be disclosed.

[0055] Although some aspects are described in the context of an apparatus, it is clear that these aspects also represent a description of a corresponding method, where a block or device corresponds to a method step or a feature of a method step. Similarly, aspects described in the context of a method step also represent a description of a corresponding block or item or feature of a corresponding apparatus.

[0056] Reference numerals list 100、102 Connector 104 Cleavage site 106 Protein backbone 108 First Reactive Interactor 110 First affinity interactor 112 Protease 114 Second affinity interactor 116 Second reactive interactor 200、202、204、206、208 Streptavidin configuration 210 Functional biotin binding site 212 Non-functional biotin binding site 300 Markers 302 dye 304、306、308、310、312 Fluorescent dyes 314 Central nucleic acid backbone 316 Oligonucleotides 400、500 Markers 402 First affinity reagent 404 Oligonucleotide of the first affinity reagent 406 Genetic structure 502 Antibody 504 Oligonucleotides for antibodies 600 Data storage device 602、704 DNA origami skeleton 604 Attachment site 700 Capture construct 702 Secondary affinity reagent 706 Affinity capture reagents 708 Capture Area

Claims

1. A connector (100, 102) for generating a biological structure (400, 500, 600, 700), comprising: Protein backbone (106), a first reactive interactant (108) disposed toward a first end of the protein backbone (106) and configured to covalently bind to a second reactive interactant (116), and a first affinity interactor (110), the affinity interactor being arranged toward the second end of the protein backbone (106) and being configured to bind to a second affinity interactor (114), wherein the protein backbone (106) comprises a cleavage site (104) between the first end and the second end of the protein backbone (106), wherein one of the first reactive interactant (108) and the first affinity interactant (110) is configured to bind to a first affinity reagent (402) comprising a corresponding second reactive interactant (116) or second affinity interactant (114), and The other of the first reactive interactant (108) and the first affinity interactant (110) is configured to bind to a label (300) or a second affinity reagent (702) including a corresponding second reactive interactant (116) or a second affinity interactant (114).

2. The linker according to claim 1, wherein the backbone (106) is specifically cleavable at the cleavage site (104) by an enzyme, cleavage light or by a temperature change.

3. The linker according to any of the preceding claims, wherein the first reactive interactant (108) and the second reactive interactant (116) are configured to form a bioconjugate.

4. The linker of any of the preceding claims, wherein the first affinity interactant (110) is one of streptavidin (200, 202, 204, 206, 208) and biotin, and the second affinity interactant (114) is the other of streptavidin (200, 202, 204, 206, 208) and biotin.

5. The linker according to claim 4, wherein the streptavidin (200, 202, 204, 206, 208) is a tetramer having one active biotin binding site (210) or a monomer having one active biotin binding site (210).

6. The linker according to any of the preceding claims, wherein the first affinity interactor (110) is streptavidin (200, 202, 204, 206, 208) and the first affinity reagent (402) is an oligonucleotide comprising biotin.

7. The linker according to any of the preceding claims, comprising the first affinity reagent (402).

8. A marker (400, 500) for analyzing a biological sample, comprising: The label (300) bound to the connector (100, 102) according to claim 7.

9. The marker according to claim 8, wherein the label (300) comprises the second reactive interactant (116) covalently bound to the first reactive interactant (108) of the linker (100, 102).

10. The marker according to any one of claims 8 or 9, comprises a third affinity reagent (502), wherein the third affinity reagent (502) comprises an oligonucleotide (504), wherein the oligonucleotide (504) is at least partially complementary to the first affinity reagent (402) of the connector (100, 102) and hybridizes with the first affinity reagent (402).

11. The marker according to claim 10, wherein the third affinity reagent (502) further comprises an antibody.

12. The marker according to any one of claims 9 to 11, wherein the marker (300) comprises: an oligonucleotide base (314) and the second reactive interactor (116); and at least one oligonucleotide-conjugated dye (200, 202, 204, 206, 208) that hybridizes to the oligonucleotide base (314) via complementary base pairing.

13. A capture construct (700) for capturing an analyte of a biological sample, comprising: a DNA origami framework (704) having at least one capture region (708), at least one affinity capture reagent (706) comprising the linker (100, 102) according to claim 7 and the second affinity reagent (702), wherein the first affinity reagent (402) of the linker (100, 102) binds the affinity capture reagent (706) to the capture region (708) of the DNA origami backbone (704), wherein the first affinity reagent (402) is a staple strand of the DNA origami backbone (704), preferably comprising the second affinity interactor (114), wherein the second affinity reagent (702) is configured to capture one of the analytes in the biological sample, and Preferably, the second affinity reagent (702) includes the second reactive interactant (116) and is covalently bound to the first reactive interactant (114) of the connector (100, 102).

14. A data storage device (600), comprising: a DNA origami backbone (602) having multiple attachment sites (604), a label (300) bound to the linker (100, 102) according to claim 7, wherein the first affinity reagent (402) is an oligonucleotide staple strand of the DNA origami backbone (602), preferably comprising the second affinity interactor (114), and The first affinity reagent (110) includes a unique oligonucleotide sequence, which is configured to bind the connector (100, 102) having the label (300) to a complementary sequence of one of the attachment sites (604).

15. A method for generating a marker (400, 500) according to any one of claims 8 to 12, comprising the following steps: combining the first affinity reagent (402) with the linker (100, 102) according to any one of claims 1 to 6, and The linker (100, 102) is combined with the label (300).

16. The method according to claim 15, comprising the steps of: The first affinity reagent (402) is combined with the third affinity reagent (502).

17. A method for producing a capture construct (700) according to claim 13 or for producing a data storage device (600) according to claim 14, comprising the following steps: combining the first affinity reagent (402) with the linker (100, 102) according to any one of claims 1 to 6, combining the linker (100, 102) with the label (300) or the second affinity reagent (702), and The first affinity reagent (402) is combined with the DNA origami backbone (602, 704).

18. A device for generating a marker (400, 500), a capture construct (700), or a data storage device (600), comprising: Liquid dispensing unit, multiple liquid containers, and A control unit configured to instruct the liquid dispensing unit to dispense the liquid containing the marker, the first affinity reagent, the second affinity reagent, the third affinity reagent or the DNA origami framework into the liquid container according to any one of the method steps of claims 15 to 17.

19. The apparatus of claim 18, wherein the liquid dispensing unit is configured to dispense the liquid using sonic droplet ejection.

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