Fully automated instrument system for biochemical assays

Abstract

This specification discloses instruments and related methods for a fully automated benchtop NULISA platform comprising an XYZ gantry, a microtiter plate stage, an incubator, a quantitative PCR module, a decontamination cleaner for microtiter plates, a microtiter plate sealer, a magnetic probe / magnetic comb assembly for sample mixing and magnetic bead extraction, a storage unit for reagents and supplies, and a controller.

Claims

1. A fully automated, compact instrument for performing a dual capture-release multiplex immunoassay, wherein the instrument is: A) It is equipped with a benchtop housing, and the benchtop housing is 1) Controller, 2) A robotic gantry capable of moving with 3 degrees of freedom. 3) A hotel capable of holding a multi-container carrier plate accessible by the user and the robot gantry, 4) A controlled stand that provides multiple multi-container carrier plate positions, the stand being movable along the Y-axis (back and forth within the housing of the equipment), 5) A mixer that operates as a magnetic bead processor and has multiple multi-container carrier plate platforms arranged vertically on top of each other. 6) A washing device for washing multi-container type carrier plates, and 7) Leader A device equipped with these features.

2. A compact instrument designed to perform a multi-step, highly sensitive immunoassay including a sequential capture / release process, wherein the instrument A) It is equipped with a benchtop housing having a touchscreen display, and the benchtop housing is 1) Controller, 2) A robotic gantry capable of moving with 3 degrees of freedom. 3) A hotel capable of holding a multi-container carrier plate accessible by the user and the robot gantry, 4) A controlled stand that provides multiple multi-container carrier plate positions, the stand being movable along the Y-axis (back and forth within the housing of the equipment), 5) A mixer that operates as a magnetic bead processor and has multiple multi-container carrier plate platforms arranged vertically on top of each other. 6) Washer for cleaning multi-container type carrier plates, 7) an incubator for incubating and sealing at least one multi-container carrier plate, 8) Leader A device equipped with these features.

3. A fully automated, high-throughput precision proteomics instrument for ultra-sensitive analysis across a wide range of multiplex levels to support extensive biomarker profiling and translation of validated biomarkers, wherein the high-throughput precision proteomics instrument is A) A benchtop housing comprising a touchscreen display, a user-accessible compartment for bulk reagents, and a user-accessible compartment for a set of bays, wherein the benchtop housing is 1) Controller, 2) A controlled robotic gantry having an end effector, comprising a pipette, a multi-container carrier plate gripper, a laser position sensor, and a barcode scanner, wherein the end effector is capable of moving with three degrees of freedom in the X, Y, and Z axes. 3) A hotel equipped with a set of multi-container carrier plate bays for housing and holding at least one universal reagent cartridge, and a consumable carrier. 4) A controlled stand that provides multiple multi-container carrier plate positions, the stand being movable along the Y-axis (back and forth within the housing of the equipment), 5) A controlled mixer that operates as a magnetic bead processor and has multiple multi-container carrier plate platforms arranged vertically on top of each other. 6) A controlled washer for cleaning multi-container carrier plates. 7) Incubator for incubating and sealing at least one multi-container carrier plate, 8) Bulk fluid stations including multiple containers, and 9) Controlled Leader A high-throughput, precision proteomics instrument equipped with these features.

4. A benchtop instrument for automatically performing multiple multiplexing oligonucleotide-conjugated antibody proximity ligation assays in parallel on multiple biological samples, wherein the benchtop instrument is: A) A benchtop housing comprising a touchscreen display, a user-accessible compartment for bulk reagents, and a user-accessible compartment for a set of bays, wherein the benchtop housing is 1) Controller, 2) A robotic gantry having an end effector, comprising a pipette, a multi-container carrier plate gripper, a laser position sensor, and a barcode scanner, wherein the end effector is capable of moving with three degrees of freedom in the X, Y, and Z axes. 3) A hotel equipped with a set of multi-container carrier plate bays for housing and holding at least one universal reagent cartridge, and a consumable carrier. 4) A stand that provides multiple multi-container type carrier plate positions, the stand being movable along the Y axis (back and forth within the housing of the equipment), 5) A mixer that operates as a magnetic bead processor and has multiple multi-container carrier plate platforms arranged vertically on top of each other. 6) Washer for cleaning multi-container type carrier plates, 7) Leader A benchtop appliance equipped with these features.

5. At least one of the multi-container carrier plates includes a pair of binding portions pre-selected to bind to a specific analyte, and the pair of binding portions is (i) A first portion comprising a first antibody or first antibody fragment pre-selected to bind to a specific analyte, a first nucleic acid target label having a first identity specific to the analyte, and a first nucleic acid tag, (ii) The second portion of the binding pair comprises a second antibody or second antibody fragment pre-selected to bind to the same specific analyte as the first antibody or first antibody fragment of the first portion of the binding pair, a second nucleic acid target label having a second identity specific to the analyte, and a second nucleic acid tag. The apparatus and / or method according to any one of claims 1 to 4, including the apparatus and / or method described in any one of claims 1 to 4.

6. A benchtop automated instrument for performing biochemical assays on biological samples containing multiple analytes, wherein the benchtop automated instrument is a) Controller and b) A controlled gantry, c) A controlled stand, d) A set of five multi-container carrier plates, e) A controlled quantitative PCR unit, f) A controlled plate washer, g) A controlled plate sealer, h) A controlled plate incubator, i) A controlled mixer equipped with a paramagnetic bead extractor with a magnetic pole / magnetic comb subassembly, j) A hotel with gantry access, 1) A plurality of first parts comprising a first antibody or capture antibody fragment conjugated with a first nucleic acid tag that hybridizes with a first nucleic acid target label but is not covalently bound to the first antibody or first antibody fragment, 2) A plurality of second parts comprising a second antibody or second antibody fragment conjugated with a second nucleic acid target label that hybridizes with a second nucleic acid target tag but is not covalently bound to the second antibody or second antibody fragment, 3) The first substrate of the first set, 4) The second substrate of the second set, 5) A set of buffers, 6) Labeling of multiple double-stranded nucleic acid samples including 5' overhangs and 3' overhangs, 7) Frame with sealing film Hotels equipped with, k) A controller comprising a processor and a non-temporary machine-readable storage medium containing instructions executable by the processor to bring about controlled operation of a unit within the device, wherein the operation controlled by the controller is 1) In the first multi-container carrier plate, i) A portion of at least one sample, ii) A portion of the plurality of first parts, and iii) A portion of the plurality of second parts To form a first solution by mixing, 2) Incubating the first solution to form an immunocomplex comprising one of the first parts from a portion of the plurality of first parts, one of the second parts from a portion of the plurality of second parts, and the analyte in at least one sample. 3) The following, i) A portion of the first substrate of the first set for capturing at least a portion of the immune complex from the first solution in the first multi-container carrier plate, wherein the first solution in the first multi-container carrier plate is incubated, and the first set of paramagnetic beads is transferred by the mixer to the second multi-container carrier plate to form a second solution, thereby releasing at least a portion of the immune complex and removing the first substrate of the first set by the mixer, ii) A portion of the second substrate of the second set for capturing at least a portion of the immune complex from the second solution in the second multi-container carrier plate, wherein the second solution is incubated and the second substrate of the second set is removed by the mixer to form a third solution in the third multi-container plate. By introducing the aforementioned immune complex sequential capture and release mechanism, contaminants are removed and sensitivity is improved. 4) Hybridize one of the overhangs with the first nucleic acid target label associated with one of the first antibodies of at least a portion of the immune complex, and hybridize the other overhangs with the second nucleic acid target label conjugated with one of the second antibodies of at least a portion of the immune complex, thereby completing proximity-based ligation between the first nucleic acid target label and the nucleic acid sample label and between the second nucleic acid target label and the nucleic acid sample label, thereby conjugating the second antibody or the second antibody fragment but the first antibody or By generating at least one multiplex analyte-specific reporter that is not covalently bound to the first antibody fragment, at least a portion of the plurality of double-stranded nucleic acid sample labels is introduced into the third solution to form at least one multiplex analyte-specific reporter, which includes the first nucleic acid target label associated with one of the first antibodies in at least one of the immune complexes, and at least one of the second nucleic acid target labels conjugated with one of the second antibodies in at least one of the immune complexes, as well as a sample-specific identity barcode. 5) Collecting at least one multiplex analyte-specific reporter in a fourth multi-container carrier plate, and then pooling the multiplex analyte-specific reporters to form a pooled sample for detection of the analyte at the atom level sensitivity by NGS. Including controllers and A benchtop automated machine equipped with these features.

7. A benchtop automated instrument for performing biochemical assays on biological samples containing multiple analytes, wherein the benchtop automated instrument is a) Controller and b) A controlled gantry, c) A controlled stand, d) A set of five multi-container carrier plates, e) A controlled quantitative PCR unit, f) A controlled plate washer, g) A controlled plate sealer, h) A controlled plate incubator, i) A controlled mixer equipped with a paramagnetic bead extractor with a magnetic pole / magnetic comb subassembly, j) A hotel with gantry access, 1) A plurality of first parts comprising a first antibody or capture antibody fragment conjugated with a first nucleic acid tag that hybridizes with a first nucleic acid target label but is not covalently bound to the first antibody or first antibody fragment, 2) A plurality of second parts comprising a second antibody or second antibody fragment conjugated with a second nucleic acid target label that hybridizes with a second nucleic acid target tag but is not covalently bound to the second antibody or second antibody fragment, 3) The first substrate of the first set, 4) The second substrate of the second set, 5) A set of buffers, 6) Labeling of multiple double-stranded nucleic acid samples including 5' overhangs and 3' overhangs, 7) Frame with sealing film Hotels equipped with, k) A controller for controlling the unit operation of the equipment, comprising a non-temporary computer-readable storage medium having an executable file, wherein the executable file is executable by the equipment controller to cause the unit operation of the equipment to perform an action, and the operation of the equipment is 1) In the first multi-container carrier plate, i) A portion of at least one sample, ii) A portion of the plurality of first parts, iii) A portion of the plurality of second parts To form a first solution by mixing, 2) Incubating the first solution to form an immunocomplex comprising one of the first parts from a portion of the plurality of first parts, one of the second parts from a portion of the plurality of second parts, and the analyte in at least one sample. 3) The following, i) A portion of the first substrate of the first set for capturing at least a portion of the immune complex from the first solution in the first multi-container carrier plate, wherein the first solution in the first multi-container carrier plate is incubated, and the first set of paramagnetic beads is transferred by the mixer to the second multi-container carrier plate to form a second solution, thereby releasing at least a portion of the immune complex and removing the first substrate of the first set by the mixer, ii) A portion of the second substrate of the second set for capturing at least a portion of the immune complex from the second solution in the second multi-container carrier plate, wherein the second solution is incubated and the second substrate of the second set is removed by the mixer to form a third solution in the third multi-container carrier plate. By introducing the aforementioned immune complex sequential capture and release mechanism, contaminants are removed and sensitivity is improved. 4) Hybridize one of the overhangs with the first nucleic acid target label associated with one of the first antibodies of at least a portion of the immune complex, and hybridize the other overhangs with the second nucleic acid target label conjugated with one of the second antibodies of at least a portion of the immune complex, thereby completing proximity-based ligation between the first nucleic acid target label and the nucleic acid sample label and between the second nucleic acid target label and the nucleic acid sample label, thereby conjugating the second antibody or the second antibody fragment but the first antibody or By generating at least one multiplex analyte-specific reporter that is not covalently bound to the first antibody fragment, at least a portion of the plurality of double-stranded nucleic acid sample labels is introduced into the third solution to form at least one multiplex analyte-specific reporter, which includes the first nucleic acid target label associated with one of the first antibodies in at least one of the immune complexes, and at least one of the second nucleic acid target labels conjugated with one of the second antibodies in at least one of the immune complexes, as well as a sample-specific identity barcode. 5) Collecting at least one multiplex analyte-specific reporter in a fourth multi-container carrier plate, and then pooling the multiplex analyte-specific reporters to form a pooled sample for detection of the analyte at the atom level sensitivity by NGS. Including controllers and A benchtop automated machine equipped with these features.

8. A benchtop automated instrument for performing a biochemical assay to detect analytes in at least one first biological sample and one second biological sample, a) Gantry and, b) Stands and, c) A multi-container carrier plate comprising a first plate, a second plate, a third plate, and a fourth plate, d) Quantitative PCR units, e) Plate washer and, f) Plate sealer, g) Plate incubator and, h) A mixer equipped with a paramagnetic bead extractor with a magnetic pole / magnetic comb subassembly, i) It is a hotel, 1) A plurality of first parts comprising a first antibody or capture antibody fragment conjugated with a first nucleic acid tag that hybridizes with a first nucleic acid target label but is not covalently bound to the first antibody or first antibody fragment, 2) A plurality of second parts comprising a second antibody or second antibody fragment conjugated with a second nucleic acid target label that hybridizes with a second nucleic acid target tag but is not covalently bound to the second antibody or second antibody fragment, 3) The first substrate of the first set, 4) The second substrate of the second set, 5) A set of buffers, 6) Labeling of multiple double-stranded nucleic acid samples including 5' overhangs and 3' overhangs, 7) Frame with sealing film Hotels equipped with, j) A controller comprising a processor and a non-temporary machine-readable storage medium containing instructions executable by the processor to bring about controlled operation of a unit within the device, wherein the operation controlled by the controller is 1) In the first multi-container carrier plate, for each of the first assay reaction containing the first biological sample and the second assay reaction containing the second biological sample, i) A portion of at least one sample, ii) A portion of the plurality of first parts, and iii) A portion of the plurality of second parts By mixing, a first solution is formed. 2) Incubating the first solution of the first assay reaction and the second assay reaction to form an immunocomplex having the analyte. 3) The following, i) A portion of the first substrate of the first set for each of the first and second assay reactions, for capturing at least a portion of the immune complex from the first solution in the first multi-container carrier plate, wherein the first solution in the first multi-container carrier plate is incubated, and the first set of paramagnetic beads is transferred by the mixer to the second multi-container carrier plate to form a second solution, releasing at least a portion of the immune complex, and the first substrate of the first set is removed by the mixer, ii) A portion of the second substrate of the second set for each of the first and second assay reactions, for capturing at least a portion of the immune complex from the second solution in the second multi-container carrier plate, wherein the second solution is incubated and the second substrate of the second set is removed by the mixer to form a third solution in the third multi-container carrier plate. By introducing this, a dual capture and release mechanism of the immune complex is performed in both the first and second assay reactions, thereby removing contaminants and improving sensitivity. 4) Hybridize one of the overhangs with the first nucleic acid target label associated with at least a portion of the first antibody of the immune complex, and hybridize the other overhangs with the second nucleic acid target label conjugated with at least a portion of the second antibody of the immune complex, thereby completing proximity-based ligation between the first nucleic acid target label and the nucleic acid sample label and between the second nucleic acid target label and the nucleic acid sample label, thereby conjugating to the second antibody or second antibody fragment but not covalently bound to the first antibody or first antibody fragment By generating at least one multiplex analyte-specific reporter, at least a portion of the plurality of double-stranded nucleic acid sample labels is introduced into the third solution for each of the first and second assay reactions to form at least one multiplex analyte-specific reporter, which includes the first nucleic acid target label associated with one of the first antibodies in at least one of the immune complexes, and at least one second nucleic acid target label conjugated with one of the second antibodies in at least one of the immune complexes, as well as a sample-specific identity barcode. 5) Collect the at least one multiplex analyte-specific reporter in a fourth multi-container carrier plate, and then pool the multiplex analyte-specific reporter from the first assay reaction together with the multiplex analyte-specific reporter from the second assay reaction to form a pooled sample for detection of the analyte at the atomol level by NGS. Including controllers and Benchtop automated equipment equipped with this feature.

9. A method for performing multiple multiplexing oligonucleotide-conjugated antibody proximity ligation assays in parallel and automatically on multiple biological samples, A) A step of forming multiple immunocomplex-forming solutions by introducing a portion of multiple multiplex-binding subgroups into multiple biological samples in parallel in a first plate, wherein the multiplex-binding subgroups of the multiple multiplex-binding subgroups include different binding subgroups that have been pre-selected to bind to different specific analytes, i) a first portion of the binding portion pair, comprising a first antibody or first antibody fragment pre-selected to bind to a specific analyte, a first nucleic acid target label having a first identity specific to the analyte, and a first nucleic acid tag, and ii) A step comprising: a second portion of the binding portion pair comprising: a second antibody or second antibody fragment pre-selected to bind to the same specific analyte as the first antibody or first antibody fragment of the first portion of the binding portion pair; a second nucleic acid target label having a second identity that is analyte-specific to the specific analyte; and a second nucleic acid tag, wherein the first nucleic acid tag of the first portion of the binding portion pair is pre-selected to bind to a first substrate which is the same for all the different binding portion pairs in the multiplex; and the second nucleic acid tag of the second portion of the binding portion pair is pre-selected to bind to a second substrate which is the same for all the different binding portion pairs in the multiplex; B) A step of incubating the plurality of immune complex-forming solutions in parallel to form a plurality of multiplex immune complexes, C) Firstly, a step of combining a portion of the first substrate solution containing a plurality of the first substrates with the plurality of immunocomplex-forming solutions, D) Firstly, a step of enabling the first nucleic acid tag of the first portion of the binding portion pair to bind to a portion of the first substrate, E) Firstly, a step of extracting the multiple multiplex immune complexes from the multiple immune complex formation solutions in parallel via a portion of the multiple first substrates to form a multiple purified first immune complex solution in a second plate, F) A step of eluting the plurality of first substrates in parallel from the plurality of multiplex immune complexes in the plurality of first immune complex purification solutions, G) A step of removing the plurality of first substrates in parallel from the plurality of the first immunocomplex purification solutions, H) Secondly, a step of combining a portion of the second substrate solution containing a plurality of the second substrates with the plurality of first immunocomplex purification solutions in parallel, I) Secondly, a step of enabling the second nucleic acid tag of the second portion of the binding portion pair to bind to a portion of the second substrate, J) Secondly, a step of extracting the multiple multiplex immune complexes from the multiple immune complex purification solutions in parallel via a portion of the multiple second substrates to form multiple second immune complex purification solutions in a third plate, K) A step of forming a plurality of multiplex analyte-specific reporters by ligating (directly or indirectly) a plurality of first nucleic acid target labels from the first portion of the binding portion pair in parallel with the second nucleic acid target labels from the second portion of the binding portion pair, L) Thirdly, the step of extracting the plurality of multiplex immune complexes (or the plurality of multiplex analyte-specific reporters) in parallel from the plurality of second immune complex purified solutions via a portion of the plurality of second substrates to form a plurality of third immune complex purified solutions in the second plate, M) A step of further eluting the plurality of second substrates in parallel from the plurality of multiplex analyte-specific reporters in the plurality of third immunocomplex purification solutions, N) A step of replicating the multiple multiplex analyte-specific reporters, O) A step of detecting the replicated multiple multiplex analyte-specific reporters to identify (and quantify) specific analytes in the multiple biological samples. A method that includes this.

10. A method for performing multiple multiplexing oligonucleotide-conjugated antibody proximity ligation assays in parallel and automatically on multiple biological samples, A) A step of forming multiple immunocomplex-forming solutions by introducing portions of multiple multiplex-binding subgroups into multiple biological samples in parallel in a first multi-container carrier plate, wherein the multiplex-binding subgroups of the multiple multiplex-binding subgroups include different binding subgroups that have been pre-selected to bind to different specific analytes, i) a first portion of the binding portion pair, comprising a first antibody or first antibody fragment pre-selected to bind to a specific analyte, a first nucleic acid target label having a first identity specific to the analyte, and a first nucleic acid tag, and ii) A step comprising: a second portion of the binding portion pair comprising: a second antibody or second antibody fragment pre-selected to bind to the same specific analyte as the first antibody or first antibody fragment of the first portion of the binding portion pair; a second nucleic acid target label having a second identity that is analyte-specific to the specific analyte; and a second nucleic acid tag, wherein the first nucleic acid tag of the first portion of the binding portion pair is pre-selected to bind to a first substrate which is the same for all the different binding portion pairs in the multiplex; and the second nucleic acid tag of the second portion of the binding portion pair is pre-selected to bind to a second substrate which is the same for all the different binding portion pairs in the multiplex; B) A step of incubating the plurality of immune complex formation solutions in parallel in a first multi-container carrier plate to form a plurality of multiplex immune complexes, C) Firstly, a step of combining a portion of a first substrate solution containing a plurality of the first substrates with the plurality of immunocomplex formation solutions in a first multi-container carrier plate, D) Firstly, a step of enabling the first nucleic acid tag of the first portion of the binding portion pair to bind to a portion of the first substrate in a first multi-container carrier plate, E) Firstly, a step of extracting the multiple multiplex immune complexes from a first multi-container carrier plate in parallel via a portion of the multiple first substrates, and depositing them onto a second multi-container carrier plate, F) A step of washing the plurality of multiplex immune complexes in the second multi-container carrier plate, G) Secondly, the process involves extracting the multiple multiplex immune complexes from the second multi-container carrier plate and depositing them onto a third multi-container carrier plate. H) A step of eluting the plurality of first substrates in parallel from the plurality of multiplex immune complexes in the third multi-container carrier plate, I) A step of removing the plurality of first substrates in parallel from the third multi-container carrier plate, J) Secondly, a step of combining a portion of the second substrate solution containing a plurality of the second substrates and the plurality of multiplex immune complexes in parallel in a third multi-container carrier plate, K) Secondly, a step of enabling the second nucleic acid tag of the second portion of the binding portion pair to bind to a portion of the second substrate in the third multi-container carrier plate, L) Thirdly, the process involves extracting the multiple multiplex immune complexes from the third multi-container carrier plate in parallel via a portion of the multiple second substrates, and depositing them onto the second multi-container carrier plate. M) Ligate a plurality of first nucleic acid target labels from the first portion of the binding portion pair (directly or indirectly) on the plurality of multiplex immunocomplexes in parallel with the second nucleic acid target labels from the second portion of the binding portion pair, thereby forming a plurality of multiplex analyte-specific reporters in the second multi-container carrier plate. N) Fourth, the process involves extracting the multiple multiplex immunocomplexes having analyte-specific reporters from the second multi-container carrier plate in parallel via a portion of the multiple second substrates, and depositing them onto the third multi-container carrier plate. O) A step of eluting the plurality of second substrates in parallel from the plurality of multiplex immune complexes having analyte-specific reporters in the third multi-container carrier plate, P) A step of replicating the multiple multiplex analyte-specific reporters, Q) A step of detecting the replicated multiple multiplex analyte-specific reporters to identify (and quantify) the presence of a specific analyte in the multiple biological samples. A method that includes this.

11. A method for performing multiple multiplexing oligonucleotide-conjugated antibody proximity ligation assays in parallel and automatically on multiple biological samples, A) A step of forming multiple immunocomplex-forming solutions by introducing portions of multiple multiplex-binding subgroups into multiple biological samples in parallel in a first multi-container carrier plate, wherein the multiplex-binding subgroups of the multiple multiplex-binding subgroups include different binding subgroups that have been pre-selected to bind to different specific analytes, i) a first portion of the binding portion pair, comprising a first antibody or first antibody fragment pre-selected to bind to a specific analyte, a first nucleic acid target label having a first identity specific to the analyte, and a first nucleic acid tag, and ii) A step comprising: a second portion of the binding portion pair comprising: a second antibody or second antibody fragment pre-selected to bind to the same specific analyte as the first antibody or first antibody fragment of the first portion of the binding portion pair; a second nucleic acid target label having a second identity that is analyte-specific to the specific analyte; and a second nucleic acid tag, wherein the first nucleic acid tag of the first portion of the binding portion pair is pre-selected to bind to a first substrate which is the same for all the different binding portion pairs in the multiplex; and the second nucleic acid tag of the second portion of the binding portion pair is pre-selected to bind to a second substrate which is the same for all the different binding portion pairs in the multiplex; B) A step of incubating the plurality of immune complex formation solutions in parallel in a first multi-container carrier plate to form a plurality of multiplex immune complexes, C) Firstly, a step of combining a portion of a first substrate solution containing a plurality of the first substrates with the plurality of immunocomplex formation solutions in a first multi-container carrier plate, D) Firstly, a step of enabling the first nucleic acid tag of the first portion of the binding portion pair to bind to a portion of the first substrate in a first multi-container carrier plate, E) Firstly, a step of extracting the multiple multiplex immune complexes from a first multi-container carrier plate in parallel via a portion of the multiple first substrates, and depositing them onto a second multi-container carrier plate, F) A step of washing the plurality of multiplex immune complexes in the second multi-container carrier plate, G) Secondly, the process involves extracting the multiple multiplex immune complexes from the second multi-container carrier plate and depositing them onto a third multi-container carrier plate. H) A step of eluting the plurality of first substrates in parallel from the plurality of multiplex immune complexes in the third multi-container carrier plate, I) A step of removing the plurality of first substrates in parallel from the third multi-container carrier plate, J) Secondly, a step of combining a portion of the second substrate solution containing a plurality of the second substrates and the plurality of multiplex immune complexes in parallel in the third multi-container carrier plate, K) Secondly, a step of enabling the second nucleic acid tag of the second portion of the binding portion pair to bind to a portion of the second substrate in the third multi-container carrier plate, L) Thirdly, the process involves extracting the multiple multiplex immune complexes from the third multi-container carrier plate in parallel via a portion of the multiple second substrates, and depositing them onto a fourth multi-container carrier plate. M) Ligate a plurality of first nucleic acid target labels from the first portion of the binding portion pair (directly or indirectly) on the plurality of multiplex immunocomplexes in parallel with the second nucleic acid target labels from the second portion of the binding portion pair, thereby forming a plurality of multiplex analyte-specific reporters in the fourth multi-container carrier plate. N) Fourth, the process involves extracting the multiple multiplex immunocomplexes having analyte-specific reporters from the fourth multi-container carrier plate in parallel via a portion of the multiple second substrates, and depositing them onto the third multi-container carrier plate. O) A step of eluting the plurality of second substrates in parallel from the plurality of multiplex immune complexes having analyte-specific reporters in the third multi-container carrier plate, P) A step of replicating the multiple multiplex analyte-specific reporters, Q) A step of detecting the replicated multiple multiplex analyte-specific reporters to identify (and quantify) the presence of a specific analyte in the multiple biological samples. A method that includes this.

12. A method for detecting at least one analyte by performing a biochemical assay on a biological sample on a benchtop automated system, 1) A step of transferring at least one sample from a multi-container sample carrier plate to a first multi-container sample carrier plate, 2) A step of diluting the sample in the first multi-container carrier plate, 3) In the second multi-container carrier plate, i) A portion of at least one sample, ii) A plurality of first parts comprising a first antibody or first antibody fragment pre-selected to bind to a specific analyte, a first nucleic acid target label having a first identity specific to the specific analyte, and a first nucleic acid tag, and iii) A plurality of portions of the second part, each comprising: a second antibody or second antibody fragment pre-selected to bind to the same specific analyte as the first antibody or first antibody fragment of the first portion of the binding portion pair; a second nucleic acid target label having a second identity specific to the analyte; and a second nucleic acid tag. A step of forming a first solution by mixing, 4) A step of incubating the first solution to form an immunocomplex containing one of the first parts from a portion of the plurality of first parts, one of the second parts from a portion of the plurality of second parts, and the analyte in at least one sample, 5) The following, i) Adding a portion of the first substrate to capture at least a portion of the immune complex from the first solution onto the second multi-container carrier plate, ii) Incubating the first solution in the second multi-container carrier plate to immobilize the immunocomplex on the surface of the first substrate, iii) Transferring the first substrate to a third multi-container carrier plate using a mixer to form a second solution having an elution buffer, iv) Wash the first substrate to remove unbound impurities. v) Incubating the second solution in a third multi-container carrier plate to release at least a portion of the immune complex, vi) Removing the first substrate with the mixer, vii) Adding a portion of the second substrate to the second solution in the third multi-container carrier plate, viiii) Incubating the second solution in the second multi-container carrier plate to immobilize the immune complex on the surface of the second substrate, ix) Transferring the second substrate to a third multi-container carrier plate using the mixer to form a third solution. x) Washing the second substrate to remove unbound impurities. This involves a process to remove contaminants and improve sensitivity by performing a dual capture and release mechanism of the immune complex, 6) A step of adding a ligation reagent to the third solution in the fourth multi-container carrier plate, 7) Hybridize one of the overhangs with the first nucleic acid target label associated with one of the first portions of at least a portion of the immune complex, and hybridize the other overhang with the second nucleic acid target label conjugated with one of the second portions of at least a portion of the immune complex, thereby completing proximity-based ligation between the first nucleic acid target label and the nucleic acid sample label and between the second nucleic acid target label and the nucleic acid sample label, which is conjugated to the second antibody or second antibody fragment but is associated with the first antibody or first antibody fragment. The process involves generating at least one unbound multiplex analyte-specific reporter, thereby introducing at least a portion of the plurality of double-stranded nucleic acid sample labels into the third solution in the fourth multi-container carrier plate to form at least one multiplex analyte-specific reporter comprising a first nucleic acid target label associated with one of the first portions of at least one of the immune complexes, and at least one second nucleic acid target label conjugated with one of the second portions of at least one of the immune complexes, as well as a sample-specific identity barcode. 8) A step of collecting at least one multiplex analyte-specific reporter into the first multi-container carrier plate, and pooling the solution of the multiplex analyte-specific reporter into the first multi-container carrier plate, 9) For detection of the analyte by NGS with atmolar-level sensitivity, the pooled solution of the multiplex analyte-specific reporter is transferred from the first plate to a qPCR plate. A method that includes this.

13. A rapid and compact instrument for automatically performing multiple multiplexing oligonucleotide-conjugated antibody proximity ligation assays on multiple biological samples, wherein the instrument is: a) A target kit, wherein the target kit is It includes a plurality of binding subgroups that are pre-selected to bind to a specific analyte, wherein the binding subgroups are i) A first portion comprising a first antibody or first antibody fragment pre-selected to bind to a specific analyte, a first nucleic acid target label having a first identity specific to the analyte, and a first nucleic acid tag, and ii) A second portion comprising a second antibody or second antibody fragment pre-selected to bind to the same specific analyte as the first antibody or first antibody fragment of the first portion of the binding portion pair, a second nucleic acid target label having a second identity specific to the analyte, and a second nucleic acid tag. A target kit including, b) A gantry comprising an end effector having a pipette and a plate gripper, an incubator, and a substrate extractor / mixer for performing double capture-release immunocomplex purification using a portion of the plurality of binding subgroups. A device equipped with these features.

14. A benchtop automated biochemical assay instrument for preparing multiplex analyte-specific reporters in a sample, a) A set of multi-container carrier plates, each having multiple wells, b) A target kit comprising multiple wells, 1) At least one first well among the plurality of wells in the target kit comprises a first portion containing a first antibody or first antibody fragment that specifically binds to the analyte, a first nucleic acid tag conjugated to the first antibody or first antibody fragment, and a first nucleic acid target label that hybridizes with the first nucleic acid tag, 2) A benchtop automated biochemical assay instrument wherein at least one second well among the plurality of wells comprises a second portion containing a second antibody or second antibody fragment that specifically binds to the analyte, a second nucleic acid label conjugated to the second antibody or second antibody fragment, and a second nucleic acid tag that hybridizes with the second nucleic acid label.

15. A rapid and compact instrument for automatically performing multiple multiplexing oligonucleotide-conjugated antibody proximity ligation assays on multiple biological samples, wherein the instrument is: a) A target kit, wherein the target kit is It includes a plurality of binding subgroups that are pre-selected to bind to a specific analyte, wherein the binding subgroups are i) A first portion comprising a first antibody or first antibody fragment pre-selected to bind to a specific analyte, a first nucleic acid target label having a first identity specific to the analyte, and a first nucleic acid tag, ii) A second portion comprising a second antibody or second antibody fragment pre-selected to bind to the same specific analyte as the first antibody or first antibody fragment of the first portion of the binding portion pair, a second nucleic acid target label having a second identity specific to the analyte, and a second nucleic acid tag. A target kit including, b) A detection kit, i) A first substrate solution containing a plurality of first substrates, ii) A second substrate solution containing a plurality of second substrates, and iii) Ligation reagents and optional nucleic acid sample-specific labeling A detection kit including, c) A microtiter sample plate containing at least 96 wells, d) Purification of dual capture-release immunocomplexes using a portion of the plurality of binding subgroups and the first and second substrate solutions, and additionally using the ligation reagent to ligate (directly or indirectly) a portion of the first nucleic acid target label of the plurality of binding subgroups with a portion of the second nucleic acid target label of the plurality of binding subgroups, thereby forming a plurality of analyte-specific reporters, comprising an end effector having petta and plate grippers, an incubator, and a gantry equipped with a substrate extractor / mixer, A device equipped with these features.

16. A benchtop instrument for automatically performing multiple multiplexing oligonucleotide-conjugated antibody proximity ligation assays in parallel on multiple biological samples, wherein the benchtop instrument is: 1) A multi-container carrier plate containing multiple biological samples, 2) A hotel, and the said hotel is a) A target kit, wherein the target kit is The system comprises multiple multiplex binding pairs, each containing different binding pairs pre-selected to bind to different specific analytes, wherein the binding pairs are i) a first portion of the binding portion pair, comprising a first antibody or first antibody fragment pre-selected to bind to a specific analyte, a first nucleic acid target label having a first identity specific to the analyte, and a first nucleic acid tag, and ii) A target kit comprising a second portion of the binding portion pair, the first portion of the binding portion pair comprising a second antibody or second antibody fragment pre-selected to bind to the same specific analyte as the first antibody or first antibody fragment of the first portion of the binding portion pair, a second nucleic acid target label having a second identity that is analyte-specific to the specific analyte, and a second nucleic acid tag, wherein the first nucleic acid tag of the first portion of the binding portion pair is pre-selected to bind to a first substrate that is the same for all the different binding portion pairs in the multiplex, and the second nucleic acid tag of the second portion of the binding portion pair is pre-selected to bind to a second substrate that is the same for all the different binding portion pairs in the multiplex, b) A detection kit, i) A first substrate solution containing a plurality of first substrates, and ii) A second substrate solution containing multiple second substrates Detection kits including A hotel equipped with, 3) A gantry equipped with an end effector having a pipette and a plate gripper, 4) Incubator and, 5) Substrate extractor / mixer, 6) qPCR and, 7) A first plate containing multiple first wells, 8) A second plate containing multiple second wells, 9) A third plate containing multiple third wells, 10) A controller, a) The controller controls the gantry to introduce a portion of the multiple multiplex binding fractions from the target kit into a portion of the multiple first wells in the first plate in parallel with a portion of the multiple biological samples, thereby forming a plurality of immunocomplex-forming solutions in a portion of the multiple first wells in the first plate. b) The controller controls the gantry and the incubator to incubate the plurality of immune complex formation solutions in parallel in portions of the plurality of first wells in the first plate, thereby forming the plurality of multiplex immune complexes. c) The controller controls the gantry to combine a portion of the first substrate solution containing the plurality of first substrates from the detection kit with the plurality of immunocomplex formation solutions in a portion of the plurality of first wells in the first plate, d) The controller controls the gantry and the incubator to enable the first nucleic acid tag of the first portion of the multiplex binding subgroup to bind to a portion of the first substrate in a portion of the plurality of first wells in the first plate. e) The controller controls the substrate extractor to extract the multiple multiplex immune complexes from the multiple immune complex formation solutions via portions of the multiple first substrates in parallel, thereby forming a plurality of purified first immune complex solutions in at least a portion of the plurality of second wells in the second plate, and the portions of the plurality of first substrates are eluted from the plurality of multiple multiplex immune complexes in the plurality of purified first immune complex solutions. f) The controller controls the substrate extractor to extract portions of the plurality of first substrates from the plurality of first immunocomplex purified solutions in parallel. g) The controller controls the gantry to combine a portion of the second substrate solution containing the plurality of second substrates from the detection kit with the plurality of first immunocomplex purified solutions in a portion of the plurality of second wells in the second plate, h) The controller controls the gantry and the incubator to enable a portion of the second nucleic acid tag of the second portion of the binding portion to a portion of the second substrate in a portion of the plurality of first immunocomplex purified solution in a portion of the plurality of second wells in the second plate, i) The controller controls the substrate extractor to simultaneously extract portions of the second substrate in portions of the first immunocomplex purified solution in portions of the second wells in the second plate via portions of the second substrate, thereby forming multiple multiplex analyte-specific reporters in portions of the third wells in the third plate, by ligating (directly or indirectly) multiple first nucleic acid target labels from the first portion of the binding portion pair in parallel with multiple second nucleic acid target labels from the second portion of the binding portion pair. j) The controller controls the substrate extractor to back-extract the plurality of multiplex analyte-specific reporters from the plurality of third wells in the third plate to the plurality of second wells in the second plate via the plurality of second substrates, thereby eluting the plurality of second substrates in parallel from the plurality of multiplex analyte-specific reporters in the plurality of second wells in the second plate. k) The controller controls the gantry and the thermocycle to replicate the multiple multiplex analyte-specific reporters, l) The controller controls the qPCR and detects the replicated multiple multiplex analyte-specific reporters, thereby identifying (and quantifying) specific analytes in the multiple biological samples. A benchtop appliance equipped with these features.

17. A method for automatically performing a dual capture-release multiplex immunoassay on multiple biological samples, below, 1) Controller, 2) A robotic gantry capable of moving with 3 degrees of freedom. 3) A hotel capable of holding a multi-container carrier plate accessible by the user and a robotic gantry. 4) A mixer that operates as a magnetic bead processor and is equipped with multiple multi-container carrier plate platforms, 5) Leader, and 6) Located within the aforementioned hotel, a) A plurality of multi-container carrier plates, each comprising a first plate containing a plurality of first wells, a second plate containing a plurality of second wells, a third plate containing a plurality of third wells, a fourth plate containing a plurality of fourth wells, and a multi-container carrier assay plate containing the plurality of biological samples. b) A multi-container carrier target kit, The system comprises multiple multiplex binding pairs, each containing different binding pairs pre-selected to bind to different specific analytes, wherein the binding pairs are i) a first portion of the binding portion pair, comprising a first antibody or first antibody fragment pre-selected to bind to a specific analyte, a first nucleic acid target label having a first identity specific to the analyte, and a first nucleic acid tag, and ii) A multi-container carrier target kit comprising a second portion of the binding portion pair, the first portion of which comprises a second antibody or second antibody fragment pre-selected to bind to the same specific analyte as the first antibody or first antibody fragment of the first portion of the binding portion pair, a second nucleic acid target label having a second identity that is analyte-specific to the specific analyte, and a second nucleic acid tag, wherein the first nucleic acid tag of the first portion of the binding portion pair is pre-selected to bind to a first substrate that is the same for all the different binding portion pairs in the multiplex, and the second nucleic acid tag of the second portion of the binding portion pair is pre-selected to bind to a second substrate that is the same for all the different binding portion pairs in the multiplex, c) A multi-container carrier detection kit, i) A first substrate solution containing a plurality of first substrates, and ii) A second substrate solution containing multiple second substrates Multi-container carrier detection kit The process includes operating equipment equipped with the following: The aforementioned device, a) By controlling the robotic gantry to introduce a portion of a plurality of multiplex binding fractions from the target kit into a portion of the plurality of first wells in the first plate, in parallel with a portion of the plurality of biological samples from the assay plate, a plurality of immunocomplex-forming solutions are formed in the portion of the plurality of first wells in the first plate. b) By controlling the robotic gantry and substrate mixer to incubate the plurality of immunocomplex formation solutions in parallel in portions of the plurality of first wells in the first plate, a plurality of multiplex immunocomplexes are formed. c) Controlling the robotic gantry to combine a portion of the first substrate solution containing the plurality of first substrates from the detection kit with the plurality of multiplex immune complexes in a portion of the plurality of first wells in the first plate, d) Control the robotic gantry and the mixer to enable a portion of the first nucleic acid tag of the first portion of the multiplex binding portion pair to bind to a portion of the first substrate in a portion of the plurality of first wells in the first plate. e) Control the mixer to extract in parallel the plurality of multiplex immune complexes bound to the plurality of first substrates from a portion of the plurality of first wells in the first plate via a portion of the plurality of first substrates, deposit them in a portion of the plurality of second wells in the second plate, and elute the plurality of first substrates from the plurality of multiplex immune complexes in a portion of the plurality of second wells in the second plate. f) Control the mixer to extract in parallel portions of the multiple first substrates from portions of the multiple multiplex immune complexes in at least a portion of the multiple second wells in the second plate, g) Controlling the robotic gantry to combine a portion of the second substrate solution containing the plurality of second substrates with the plurality of multiplex immune complexes in a portion of the plurality of second wells in the second plate, h) Control the robotic gantry and the mixer to enable a portion of the second nucleic acid tag of the second portion of the binding portion pair to bind to a portion of the second substrate in a portion of the plurality of second wells in the second plate. i) Control the mixer to parallel extract portions of the multiple multiplex immunocomplexes bound to portions of the second substrate in portions of the multiple second wells in the second plate via portions of the multiple second substrates, deposit them in portions of the multiple third wells in the third plate, and form multiple multiplex analyte-specific reporters by ligating (directly or indirectly) multiple first nucleic acid target labels from the first portion of the binding portion pair in parallel with multiple second nucleic acid target labels from the second portion of the binding portion pair. j) Control the mixer to extract the multiple multiplex analyte-specific reporters bound to the portion of the second substrate from a portion of the multiple third wells in the third plate via a portion of the multiple second substrates, and deposit them back into a portion of the multiple second wells in the second plate, thereby eluting the multiple second substrates in parallel from the multiple multiplex analyte-specific reporters in a portion of the multiple second wells in the second plate. k) Controlling the robotic gantry to transfer at least a portion of the plurality of multiple analyte-specific reporters from a portion of the plurality of second wells in the second plate to a portion of the plurality of fourth wells in the fourth plate, l) Control the robot gantry to move the fourth plate to the leader. A way of working.

18. A fully automated, compact instrument for performing a dual capture-release multiplex immunoassay, wherein the instrument is: A) It is equipped with a benchtop housing, and the benchtop housing is 1) A robotic gantry capable of moving with 3 degrees of freedom. 2) A hotel capable of holding a multi-container carrier plate accessible by the user and a robotic gantry. 3) A controlled stand that provides multiple multi-container carrier plate positions, the stand being movable along the Y-axis (back and forth within the housing of the equipment), and 4) A mixer that operates as a magnetic bead processor and has multiple multi-container carrier plate platforms arranged vertically on top of each other. A device equipped with these features.

19. The apparatus and / or method according to any one of claims 1 to 18, wherein one of the first nucleic acid tag or the second nucleic acid tag includes a nucleotide-rich sequence.

20. The apparatus and / or method according to any one of claims 1 to 19, wherein one of the first nucleic acid tag or the second nucleic acid tag comprises one or more nucleotide-rich sequences selected from poly-A, poly-T, poly-C, or poly-G sequences.

21. The apparatus and / or method according to any one of claims 1 to 20, wherein the first nucleic acid tag comprises a poly-A sequence or a poly-T sequence.

22. The apparatus and / or method according to any one of claims 1 to 21, wherein one of the first nucleic acid tag or the second nucleic acid tag includes an immobilization reagent.

23. The apparatus and / or method according to any one of claims 1 to 22, wherein one of the first nucleic acid tag or the second nucleic acid tag comprises one or more immobilization reagents from among biotin, streptavidin, EDC, DCC, NHS ester, imide ester, maleimide, haloacetyl, pyridyl disulfide, hydrazide, alkoxyamine, aryl azide, didiline, or a chemoselective ligation group.

24. The apparatus and / or method according to any one of claims 1 to 23, wherein the second nucleic acid tag is conjugated with biotin.

25. The apparatus and / or method according to any one of claims 1 to 24, wherein the first nucleic acid tag is not covalently bound to the first antibody or the first antibody fragment.

26. The apparatus and / or method according to any one of claims 1 to 25, wherein the first nucleic acid target label is not covalently bound to the first antibody or the first antibody fragment.

27. The apparatus and / or method according to any one of claims 1 to 26, wherein the second nucleic acid tag is not covalently bound to the second antibody or the second antibody fragment.

28. The apparatus and / or method according to any one of claims 1 to 27, wherein the first antibody or the first antibody fragment and the second antibody or the second antibody fragment are paired to bind to a non-overlapping epitope of the analyte to form an immune complex.

29. The first antibody or the first antibody fragment has a binding affinity of at least 10 to the analyte. －４ M is present, and the second antibody or the second antibody fragment has a binding affinity of at least 10 to the analyte. －４ The apparatus and / or method according to any one of claims 1 to 28, wherein M.

30. The apparatus and / or method according to any one of claims 1 to 29, wherein the first nucleic acid target label includes a first identity barcode.

31. The apparatus and / or method according to any one of claims 1 to 30, wherein the second nucleic acid target label includes a second identity barcode.

32. The apparatus and / or method according to any one of claims 1 to 31, wherein the apparatus further comprises a target kit.

33. The target kit comprises a plurality of wells of the multi-container carrier plate, (i) At least one first well among the plurality of wells in the target kit includes the first portion, (ii) The apparatus and / or method according to any one of claims 1 to 32, wherein at least one second well among the plurality of wells in the target kit includes the second portion.

34. The apparatus and / or method according to any one of claims 1 to 33, wherein the first well and the second well are the same wells of the multi-container carrier plate.

35. The apparatus and / or method according to any one of claims 1 to 34, wherein the first well and the second well are different wells of the multi-container carrier plate.

36. The target kit further comprises a plurality of wells of the multi-container carrier plate, (i) At least one third well of the plurality of wells of the target kit contains a first non-functional binder comprising a third antibody or a third antibody fragment that specifically binds to the analyte, (ii) The apparatus and / or method according to any one of claims 1 to 35, wherein at least one fourth well of the plurality of wells of the target kit contains a second non-functional binder comprising a fourth antibody or a fourth antibody fragment that specifically binds to the analyte.

37. The apparatus and / or method according to any one of claims 1 to 36, wherein the first non-functional binder binds to the same epitope of the analyte in the same manner as the first portion.

38. The apparatus and / or method according to any one of claims 1 to 37, wherein the third antibody or the third antibody fragment is identical to the first antibody or the first antibody fragment of the first portion.

39. The apparatus and / or method according to any one of claims 1 to 38, wherein the fourth antibody or the fourth antibody fragment is identical to the second antibody or the second antibody fragment of the second portion.

40. The apparatus and / or method according to any one of claims 1 to 39, wherein the third well and the fourth well are the same wells of the multi-container carrier plate.

41. The apparatus and / or method according to any one of claims 1 to 40, wherein the third well and the fourth well are different wells of the multi-container carrier plate.

42. The apparatus and / or method according to any one of claims 1 to 41, wherein the first well, the second well, the third well, and the fourth well are the same wells of the multi-container carrier plate.

43. The apparatus and / or method according to any one of claims 1 to 42, wherein the first well, the second well, the third well, and the fourth well are different wells of the multi-container carrier plate.

44. The apparatus and / or method according to any one of claims 1 to 43, wherein the first non-functional binder is mixed with the first portion in a predetermined proportion.

45. The apparatus and / or method according to any one of claims 1 to 44, wherein the second non-functional binder is mixed with the second portion in a predetermined proportion.

46. The apparatus and / or method according to any one of claims 1 to 45, further comprising a detection kit.

47. The detection kit comprises multiple wells of a multi-container carrier plate, (i) At least one of the plurality of wells contains a first substrate solution containing a plurality of first substrates, (ii) At least one second well among the plurality of wells contains a second substrate solution containing a plurality of second substrates, (iii) The apparatus and / or method according to any one of claims 1 to 46, wherein at least one third well among the plurality of wells contains a ligation reagent and optionally a nucleic acid sample-specific label.

48. The apparatus and / or method according to any one of claims 1 to 47, wherein the first substrate is a first paramagnetic bead.

49. The apparatus and / or method according to any one of claims 1 to 48, wherein the first paramagnetic beads are bound to a first binding sequence capable of binding to the first nucleic acid tag on the first antibody or the first antibody fragment.

50. The apparatus and / or method according to any one of claims 1 to 49, wherein the first paramagnetic beads are coated with a first binding sequence capable of binding to the first nucleic acid tag on the first antibody or the first antibody fragment.

51. The apparatus and / or method according to any one of claims 1 to 50, wherein the second substrate is a second paramagnetic bead.

52. The apparatus and / or method according to any one of claims 1 to 51, wherein the second paramagnetic bead is bound to a second binding group capable of binding to the second antibody or the second nucleic acid tag associated with the second antibody fragment.

53. The apparatus and / or method according to any one of claims 1 to 52, wherein the second paramagnetic bead is coated with a second binding group capable of binding to the second antibody or the second nucleic acid tag associated with the second antibody fragment.

54. The apparatus and / or method according to any one of claims 1 to 53, wherein the first binding sequence is pre-selected to hybridize with the first nucleic acid tag on the first antibody or the first antibody fragment.

55. The apparatus and / or method according to any one of claims 1 to 54, wherein the first bonding arrangement bonded to the first paramagnetic bead is a poly-A arrangement or a poly-T arrangement.

56. The apparatus and / or method according to any one of claims 1 to 55, wherein a poly-A sequence from the first nucleic acid tag is hybridized with a poly-T sequence from the first paramagnetic beads.

57. The apparatus and / or method according to any one of claims 1 to 56, wherein a poly-T sequence from the first nucleic acid tag is hybridized with a poly-A sequence from the first paramagnetic beads.

58. The apparatus and / or method according to any one of claims 1 to 57, wherein the second binding group bonded to the second paramagnetic bead is streptavidin or avidin.

59. The apparatus and / or method according to any one of claims 1 to 58, wherein the biotin from the second nucleic acid tag is bound to streptavidin from the second paramagnetic beads.

60. The apparatus and / or method according to any one of claims 1 to 59, wherein the biotin from the second nucleic acid tag is bound to avidin from the second paramagnetic beads.

61. The apparatus and / or method according to any one of claims 1 to 60, wherein the binding between the first nucleic acid tag and the binding sequence is orthogonal.

62. The apparatus and / or method according to any one of claims 1 to 61, wherein the bond between the second nucleic acid tag and the binding group is an orthogonal bond.

63. The apparatus and / or method according to any one of claims 1 to 62, wherein the first well, the second well, and the third well of the multi-container carrier plate of the detection kit are located in the multi-container carrier plate of the target kit.

64. The apparatus and / or method according to any one of claims 1 to 63, wherein the first well and the second well of the target kit are located in the detection kit.

65. The apparatus and / or method according to any one of claims 1 to 64, wherein the apparatus further comprises an incubator for incubating and / or sealing at least one multi-container carrier plate.

66. The apparatus and / or method according to any one of claims 1 to 65, wherein the apparatus further comprises a bulk fluid station having a plurality of containers.

67. The apparatus and / or method according to any one of claims 1 to 66, wherein the robotic gantry further comprises an end effector equipped with a pipette, a multi-container carrier plate gripper, a laser position sensor, and a barcode scanner, and the end effector is capable of moving with three degrees of freedom in the X, Y, and Z axes.

68. The apparatus and / or method according to any one of claims 1 to 67, wherein the reader includes a qPCR unit for qPCR readout.

69. The apparatus and / or method according to any one of claims 1 to 68, wherein the reader includes qPCR for preparing a pooled library to be prepared for next-generation sequencing (NGS).

70. The apparatus and / or method according to any one of claims 1 to 69, wherein the reader is capable of identifying and / or quantifying nucleic acid reporters.

71. The apparatus and / or method according to any one of claims 1 to 70, wherein the housing further comprises a touchscreen display.

72. The apparatus and / or method according to any one of claims 1 to 71, wherein the hotel comprises a set of multi-container carrier bays for housing and holding at least one universal reagent cartridge, and a consumables carrier.

73. The aforementioned hotel, a) The target kit and, b) The detection kit and, c) The at least one consumable carrier, i) PCR multi-container carrier plate, ii) Sealing film with frame, iii) Multiple pipette tips, and iv) Multiple multi-container carrier plates Includes at least one consumable carrier and The apparatus and / or method according to any one of claims 1 to 72, further comprising:

74. The apparatus and / or method according to any one of claims 1 to 73, wherein the framed sealing film has one or more perforations configured to allow the film to be easily and completely separated from the frame by tearing.

75. The apparatus and / or method according to any one of claims 1 to 74, wherein the framed sealing film seals the well when it is heat-welded to the edge of the multi-container carrier plate.

76. The apparatus and / or method according to any one of claims 1 to 75, wherein the framed sealing film includes a metal layer.

77. The apparatus and / or method according to any one of claims 1 to 76, wherein the framed sealing film is perforated by a pipette tip.

78. The apparatus and / or method according to any one of claims 1 to 77, wherein the controller comprises a processor and a non-temporary machine-readable storage medium containing instructions executable by the processor to give controlled operation of components within the apparatus.

79. The apparatus and / or method according to any one of claims 1 to 78, wherein the controller comprises a processor and a non-temporary machine-readable storage medium containing pre-programmed instructions that can be executed by the processor to perform a dual capture-release multiplex immunoassay.

80. The controller comprises a processor and a non-temporary machine-readable storage medium containing instructions executable by the processor to give controlled operation of a unit in the system, and the operation controlled by the controller is a) In the first multi-container carrier plate, i) At least one portion of the biological sample, ii) A portion of the plurality of first parts, and iii) A portion of the plurality of second parts To form a first solution by mixing, b) Incubating the first solution in the incubator to form an immunocomplex comprising one of the first parts from a portion of the plurality of first parts, one of the second parts from a portion of the plurality of second parts, and the analyte in at least one biological sample. c) Below, i) A portion of the first paramagnetic beads for capturing at least a portion of the immune complex from the first solution in the first multi-container carrier plate, wherein the first solution in the first multi-container carrier plate is incubated, the first paramagnetic beads are transferred by the extractor / mixer to a second multi-container carrier plate to form a second solution, releasing at least a portion of the immune complex, and the first paramagnetic beads are removed by the extractor / mixer. ii) A portion of the second paramagnetic beads for capturing at least a portion of the immune complex from the second solution, wherein the second solution is incubated and the second set of paramagnetic beads is removed by the extractor / mixer, thereby forming the third solution in the third multi-container carrier plate. By introducing this mechanism, a dual capture and release mechanism of the immune complex is achieved, thereby removing unbound contaminants and improving sensitivity. Apparatus and / or method according to any one of claims 1 to 79, including

81. The apparatus and / or method according to any one of claims 1 to 80, wherein the contaminant may include an unbound first portion or an unbound second portion.

82. The apparatus and / or method according to any one of claims 1 to 81, wherein the contaminant may include a non-target analyte.

83. The apparatus and / or method according to any one of claims 1 to 82, wherein the contaminant may include an unbound target analyte.

84. The operation controlled by the controller is a) Washing the first solid surface of the first substrate to remove unbound contaminants, and b) Washing the second solid surface of the second substrate to remove unbound contaminants. The apparatus and / or method according to any one of claims 1 to 83, further comprising:

85. The operation controlled by the system controller is a) Hybridizing one of the overhangs with the first nucleic acid target label associated with one of the first portions of at least a portion of the immune complex, and hybridizing the other overhangs with the second nucleic acid target label conjugated with one of the second portions of at least a portion of the immune complex, thereby completing proximity-based ligation between the first nucleic acid target label and the nucleic acid sample label and between the second nucleic acid target label and the nucleic acid sample label, thereby generating at least one nucleic acid reporter conjugated to the first portion but not covalently bonded to the second portion, thereby introducing at least a portion of the plurality of nucleic acid sample labels into the third solution to form at least one nucleic acid reporter comprising the first nucleic acid target label associated with one of the first portions of at least one of the immune complexes, and at least one second nucleic acid target label conjugated with one of the second portions of at least one of the immune complexes, and a sample-specific identity barcode, b) Collecting at least one nucleic acid reporter into a fourth multi-container carrier plate, and then pooling the nucleic acid reporters, to form a pooled sample for detection of the analyte by NGS with an atmolar level sensitivity having a wide dynamic measurement range. The apparatus and / or method according to any one of claims 1 to 84, further comprising:

86. The apparatus and / or method according to any one of claims 1 to 85, further comprising the operation controlled by the controller washing the multi-container carrier plate previously used to perform the capture and release mechanism by a multi-container carrier plate washer, and reintroducing the immune complex into the washed multi-container carrier plate.

87. The apparatus and / or method according to any one of claims 1 to 86, wherein the apparatus is pre-configured to perform a dual capture-release multiplex immunoassay.

88. The apparatus and / or method according to any one of claims 1 to 87, wherein the apparatus is an all-in-one instrument with an integrated qPCR instrument, and processes the sample assay from sample preparation to data or a pooled NGS library.

89. The apparatus and / or method according to any one of claims 1 to 88, wherein the apparatus is capable of detection at an atomographic sensitivity.

90. The apparatus and / or method according to any one of claims 1 to 89, wherein the apparatus is capable of detection with an atomographic sensitivity of at least 5.

91. The apparatus and / or method according to any one of claims 1 to 90, wherein the apparatus is capable of atomographic sensitivity detection having a wide dynamic measurement range of up to 12 logs, including an upper detection limit for one member of the target panel and a lower detection limit for one member of the target panel.

92. The apparatus and / or method according to any one of claims 1 to 91, wherein the apparatus is capable of performing a dual capture-release multiplex immunoassay in parallel on more than 15 samples.

93. The apparatus and / or method according to any one of claims 1 to 92, wherein the apparatus can complete a dual capture-release multiplex immunoassay on multiple samples from an assay sample without human intervention.

94. The apparatus and / or method according to any one of claims 1 to 93, wherein the apparatus can complete a dual capture-release multiplex immunoassay on multiple samples from an assay sample in less than four hours without human intervention.

95. The apparatus and / or method according to any one of claims 1 to 94, wherein the apparatus can complete a dual capture-release multiplex immunoassay on multiple samples from an assay sample, up to qPCR readout, in less than 6 hours without human intervention.

96. The apparatus and / or method according to any one of claims 1 to 95, wherein the apparatus automatically completes a multiplex oligonucleotide-conjugated antibody proximity ligation assay on a plurality of biological samples within 6 hours, with a low sensitivity of about 50 atmoles, from sample preparation to the purification of a dual capture-release immunocomplex and the formation of a plurality of analyte-specific reporters for a multiplex of analytes in the plurality of biological samples.

97. The apparatus and / or method according to any one of claims 1 to 96, wherein the apparatus completes the dual capture and release mechanism in less than two hours.

98. The apparatus and / or method according to any one of claims 1 to 97, wherein the apparatus includes a sufficient amount of reagents and other consumables to perform at least three cycles of the assay without the user reloading the reagents and other consumables.

99. The apparatus and / or method according to any one of claims 1 to 98, wherein the apparatus is a benchtop apparatus.

100. The apparatus and / or method according to any one of claims 1 to 99, wherein the apparatus is less than 50 inches in height (excluding the stand).

101. The apparatus and / or method according to any one of claims 1 to 100, wherein the base of the apparatus is less than 30 inches deep and less than 50 inches wide.

102. The apparatus and / or method according to any one of claims 1 to 101, wherein the apparatus is capable of high throughput.

103. The apparatus and / or method according to any one of claims 1 to 102, wherein the sample is a serum sample, a plasma sample, or a cerebrospinal fluid sample.

104. The apparatus and / or method according to any one of claims 1 to 103, wherein the sample is derived from a cell sample or a tissue sample.

105. The apparatus and / or method according to any one of claims 1 to 104, wherein the sample has more than 50 analytes.

106. The apparatus and / or method according to any one of claims 1 to 105, wherein the amount of the sample is less than 20 microliters.

107. The apparatus and / or method according to any one of claims 1 to 106, wherein the analyte is a protein or polypeptide.

108. The apparatus and / or method according to any one of claims 1 to 107, wherein the multiplex detects at least 50 analytes.

109. An automated instrument for performing a dual capture-release immunoassay procedure, which consumes five or fewer multi-container carrier plates for assay execution.

110. An automated instrument for performing a dual capture-release immunoassay procedure, which consumes two or fewer boxes of pipette tips for the execution of the assay.

111. The apparatus and / or method according to any one of claims 1 to 110, wherein the apparatus carries a solution containing the immune complex using at least one plate to capture the immune complex on a substrate, and reuses the at least one plate to carry the solution containing the immune complex and elute the immune complex from the substrate.

112. The apparatus and / or method according to any one of claims 1 to 111, wherein the apparatus uses at least one plate to carry a solution containing the immune complex and capture the immune complex on a substrate, washes the at least one plate, and reuses the at least one plate to carry a solution containing the immune complex and elute the immune complex from the substrate.

113. The apparatus and / or method according to any one of claims 1 to 112, wherein the apparatus carries a solution containing the immunocomplex bound on the first substrate by using at least one plate, and carries a solution containing the immunocomplex bound on the second substrate by reusing the at least one plate.

114. The apparatus and / or method according to any one of claims 1 to 113, wherein the apparatus carries a solution containing the immunocomplex bound on the first substrate using at least one plate to elute the first substrate from the immunocomplex, and carries a solution containing the immunocomplex bound on the second substrate using at least one plate to elute the second substrate from the immunocomplex.

115. The apparatus and / or method according to any one of claims 1 to 114, wherein the apparatus carries a solution containing the immune complex using at least one plate to elute the first substrate from the immune complex, and reuses the at least one plate to carry a solution containing the immune complex to elute the second substrate from the immune complex.

116. The apparatus and / or method according to any one of claims 1 to 115, wherein the apparatus carries a solution containing the immune complex using at least one plate to elute the first substrate from the immune complex, washes the at least one plate, and reuses the at least one plate to carry a solution containing the immune complex to elute the second substrate from the immune complex.

117. The apparatus and / or method according to any one of claims 1 to 116, wherein the apparatus carries a solution containing the immune complex using at least one plate, captures the immune complex on a second substrate, and reuses the at least one plate to carry the solution containing the immune complex and elute the second substrate from the immune complex.

118. The apparatus and / or method according to any one of claims 1 to 117, wherein the apparatus transports a solution containing the immune complex using at least one plate, then captures the immune complex on a second substrate, and after capturing the immune complex on the second substrate, reuses the at least one plate to transport the solution containing the immune complex.

119. The apparatus and / or method according to any one of claims 1 to 118, wherein the apparatus carries a solution containing the immune complex bound to the first substrate by using at least one plate, and carries the solution containing the immune complex by using at least one plate to capture the immune complex on a second substrate.

120. The apparatus and / or method according to any one of claims 1 to 119, wherein the apparatus carries a solution containing the immunocomplex bound to the first substrate by using at least one plate, and carries the solution containing the immunocomplex by reusing the at least one plate to elute the immunocomplex from the second substrate.

121. The apparatus and / or method according to any one of claims 1 to 120, wherein the apparatus carries a solution containing the immune complex bound to the first substrate by using at least one plate, carries the solution containing the immune complex by using at least one plate to capture the immune complex on a second substrate, and reuses the at least one plate to carry the solution containing the immune complex to elute the immune complex from the second substrate.

122. The apparatus and / or method according to any one of claims 1 to 121, wherein the apparatus, during the elution of the first substrate, carries a solution containing the immune complex bound to the first substrate by using at least one plate, carries the solution containing the immune complex by using at least one plate to capture the immune complex on the second substrate, and reuses the at least one plate to carry the solution containing the immune complex to elute the immune complex from the second substrate.

123. The apparatus and / or method according to any one of claims 1 to 122, wherein the apparatus transports a solution containing the immune complex using at least one plate, captures the immune complex on a second substrate, washes the at least one plate, and then, after capturing the immune complex on the second substrate, reuses the at least one plate to transport the solution containing the immune complex.

124. The apparatus and / or method according to any one of claims 1 to 123, wherein the apparatus carries a solution containing the immunocomplex bound to the first substrate by using at least one plate, washes the at least one plate, and reuses the at least one plate to carry the solution containing the immunocomplex and capture the immunocomplex on the second substrate.

125. The apparatus and / or method according to any one of claims 1 to 124, wherein the apparatus carries a solution containing the immunocomplex bound to the first substrate by using at least one plate, washes the at least one plate, and reuses the at least one plate to carry the solution containing the immunocomplex and elute the immunocomplex from the second substrate.

126. The apparatus and / or method according to any one of claims 1 to 125, wherein the apparatus carries a solution containing the immune complex bound to the first substrate by using at least one plate, washes the at least one plate, reuses the at least one plate to carry the solution containing the immune complex and capture the immune complex on the second substrate, and reuses the at least one plate to carry the solution containing the immune complex and elute the immune complex from the second substrate.

127. The apparatus and / or method according to any one of claims 1 to 126, wherein the apparatus, during the elution of the first substrate, carries a solution containing the immune complex bound to the first substrate by using at least one plate, washes the at least one plate, reuses the at least one plate to carry the solution containing the immune complex to capture the immune complex on the second substrate, and reuses the at least one plate to carry the solution containing the immune complex to elute the immune complex from the second substrate.

128. The apparatus and / or method according to any one of claims 1 to 127, wherein the apparatus uses at least one plate to carry an immune complex solution and capture the immune complex on a substrate, then washes the at least one plate and reuses the at least one plate to elute the immune complex solution from the substrate.

129. The apparatus and / or method according to any one of claims 1 to 128, wherein the apparatus holds the immunocomplex solution by using at least one plate during washing, and holds the immunocomplex solution by reusing the at least one plate during ligation to form the reporter.

130. The apparatus and / or method according to any one of claims 1 to 129, wherein the apparatus holds a solution containing the immunocomplex bound to the first substrate by using at least one plate, and holds a solution containing the immunocomplex bound to the second substrate by reusing the at least one plate.

131. The apparatus and / or method according to any one of claims 1 to 130, wherein the apparatus holds a solution containing the immunocomplex bound to the first substrate by using at least one plate during washing, and holds a solution containing the immunocomplex bound to the second substrate by reusing the at least one plate during ligation.

132. The apparatus and / or method according to any one of claims 1 to 131, wherein the apparatus holds a solution containing the immunocomplex bound to the first substrate by using at least one plate, then elutes the first substrate, and then holds a solution containing the immunocomplex bound to the second substrate by reusing the at least one plate, then elutes the second substrate.

133. The apparatus and / or method according to any one of claims 1 to 132, wherein the apparatus holds the immune complex formation solution and forms the immune complex by using at least one plate, and captures the immune complex on a substrate by using at least one plate.

134. The apparatus and / or method according to any one of claims 1 to 133, wherein the apparatus holds the immune complex formation solution and forms the immune complex using at least one plate, and captures the immune complex on a first substrate using at least one plate.

135. The apparatus and / or method according to any one of claims 1 to 134, wherein the apparatus holds the immune complex forming solution and forms the immune complex by using at least one plate, captures the immune complex on a first substrate by using at least one plate, and holds the solution containing the immune complex solution bound to the first substrate by reusing the at least one plate.

136. The apparatus and / or method according to any one of claims 1 to 135, wherein the apparatus holds the immune complex forming solution and forms the immune complex by using at least one plate, captures the immune complex on a first substrate by using at least one plate, washes the at least one plate, and holds the solution containing the immune complex solution bound to the first substrate by reusing the at least one plate.

137. The apparatus and / or method according to any one of claims 1 to 136, wherein the apparatus holds the immune complex forming solution and forms the immune complex using at least one plate, captures the immune complex on a first substrate using at least one plate, and holds the solution containing the immune complex solution bound to the first substrate and performs washing by reusing the at least one plate.

138. The apparatus and / or method according to any one of claims 1 to 137, wherein the apparatus holds the immune complex forming solution and forms the immune complex using at least one plate, captures the immune complex on a first substrate using at least one plate, washes the at least one plate, and reuses the at least one plate to hold the solution containing the immune complex solution bound to the first substrate and washes it.

139. The apparatus and / or method according to any one of claims 1 to 138, wherein the apparatus holds the immune complex solution and captures the immune complex on a substrate by using at least the same plate, and elutes the immune complex from a substate by using at least the same plate.

140. The apparatus and / or method according to any one of claims 1 to 139, wherein the apparatus holds the immune complex formation solution and forms the immune complex by using at least the same plate, and captures the immune complex on the first substrate by using at least the same plate.

141. The apparatus and / or method according to any one of claims 1 to 140, wherein the apparatus retains the immunocomplex and elutes the first substrate by using at least the same plate, and captures the immunocomplex on the first substrate by using at least the same plate.

142. The apparatus and / or method according to any one of claims 1 to 141, wherein the apparatus holds the immunocomplex solution and elutes the first substrate by using at least the same plate, and captures the immunocomplex on the second substrate by using at least the same plate.

143. The apparatus and / or method according to any one of claims 1 to 142, wherein the apparatus holds the first substrate solution and the second substrate solution by using at least the same plate.

144. The apparatus and / or method according to any one of claims 1 to 143, wherein the apparatus holds the first substrate solution by using at least the same plate, washes at least the same plate, and holds the second substrate solution by reusing the same plate.

145. The apparatus and / or method according to any one of claims 1 to 144, wherein the apparatus carries a solution containing the target analyte from one of the steps of the process by reusing at least one plate in a later step of the process.

146. The apparatus and / or method according to any one of claims 1 to 145, wherein the apparatus carries the solution containing the immunocomplex from one step of the process by reusing at least one plate in a later step of the process.

147. The apparatus and / or method according to any one of claims 1 to 146, wherein the apparatus holds a solution containing at least one immunocomplex that is a precursor to a target analyte by using at least one plate, and in a later step of the process, the solution containing the immunocomplex is transported from one step of the process by reusing the at least one plate.

148. The apparatus and / or method according to any one of claims 1 to 147, wherein the apparatus carries the sample, which means the target analyte or a solution containing the analyte captured by the immune complex, from the formation of the immune complex through various process steps to the formation of the corresponding nucleic acid analyte reporter, without any pipette tip coming into contact with the sample.

149. The apparatus and / or method according to any one of claims 1 to 148, wherein the apparatus carries the sample, which means the target analyte or a solution containing the analyte captured by the immune complex, from the formation of the immune complex through the process steps to the formation of the corresponding nucleic acid analyte reporter, without requiring any pipette tip to come into contact with the sample.

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