Diagnostic device

The diagnostic device uses a microporous phase material with electromagnetically activatable crosslinking agents to enhance the binding and detection of target analytes, addressing inefficiencies in existing diagnostic methods.

HK40135695APending Publication Date: 2026-07-31HUTANO DIAGNOSTICS LTD
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Patent Information

Application Number
HK62026123535
Authority / Receiving Office
HK · HK
Patent Type
Applications
Current Assignee / Owner
Priority Date
2023-01-09
Filing Date
2026-05-18
Publication Date
2026-07-31

AI Technical Summary

Technical Problem

Existing diagnostic devices lack efficient and reliable methods for binding and detecting target analytes using electromagnetically activatable crosslinking agents covalently bound to microporous phase materials.

Method used

A diagnostic device incorporating a microporous phase material with a capture reagent covalently bound via electromagnetically activatable functional groups, capable of binding target analytes through a covalently bound portion and a detection reagent, utilizing a first linker portion such as an aptamer, and a second linker portion for detection.

Benefits of technology

Enhances the binding and detection efficiency of target analytes by leveraging electromagnetically activatable crosslinking agents, providing a reliable and efficient diagnostic method.

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Abstract

Described is a diagnostic device, such as a lateral flow device comprising: a microporous phase material, a capture reagent covalently bound to a capture zone of said microporous phase material, wherein said capture reagent comprises a covalent binding moiety, wherein the covalent binding moiety comprises an electromagnetic radiation (EMR)–activable cross linker having an EMR -activable functional group, wherein the capture reagent is covalently bound to said capture zone via the functional group activated by electromagnetic radiation, and wherein said capture reagent is capable of binding an analyte of interest. The capture reagent may comprise (i) a first linker moiety which may be an oligonucleotide for example an aptamer, wherein the covalent binding moiety is at the 5' or 3' terminus of the linker moiety, and / or a detection reagent comprising a second linker moiety, wherein said detection reagent is capable of binding the analyte of interest.
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Description

Invention Title: Diagnostic Device This invention discloses a diagnostic device, such as a side-flow device, comprising: a microporous phase material, and a capture reagent covalently bound to a capture region of the microporous phase material, wherein the capture reagent includes a covalently bound portion comprising an electromagnetically activatable crosslinking agent having electromagnetically activatable functional groups; the capture reagent is covalently bound to the capture region via electromagnetically activated functional groups, and the capture reagent is capable of binding a target analyte. The capture reagent may include (i) a first linker portion, which may be an oligonucleotide, such as an aptamer, wherein the covalently bound portion is located at the 5' or 3' end of the linker portion; and / or a detection reagent including a second linker portion, the detection reagent being capable of binding the target analyte. Abstract

Claims

Claims1. A diagnostic device comprising: a microporous phase material, a capture reagent covalently bound to a predetermined capture zone of said microporous phase material, wherein said capture reagent comprises a covalent binding moiety, wherein the covalent binding moiety comprises a long-wave electromagnetic radiation (EMR)-activable cross linker having a long wave electromagnetic radiation (EMR) activable functional group, wherein the capture reagent is covalently bound to said capture zone via the functional group activated by electromagnetic radiation, and wherein said capture reagent is capable of binding an analyte of interest; wherein either:(i) said capture reagent further comprises a first linker moiety wherein the said covalent binding moiety is at the 5’ or 3’ terminus of the first linker moiety; and / or(ii) said diagnostic device comprises a detection reagent comprising a second linker moiety, wherein said detection reagent is capable of binding the analyte of interest; wherein each linker moiety comprises or consists of either an oligonucleotide moiety, a hydrocarbon chain, or an oligonucleotide moiety linked to a hydrocarbon chain, optionally wherein said hydrocarbon chain is a linear hydrocarbon chain.

2. The diagnostic device according to claim 1, wherein said capture reagent comprises said first linker moiety, wherein said first linker moiety is a first oligonucleotide moiety and wherein the said covalent binding moiety is at the 5’ or 3’ terminus of the first oligonucleotide moiety.

3. The diagnostic device according to claim 1 or claim 2, wherein said capture reagent comprises an antibody molecule with binding specificity for the analyte of interest.

4. The diagnostic device according to claim 3, wherein the covalent binding moiety is attached to a Fc region of the antibody molecule.

5. The diagnostic device according to claim 2, wherein the capture reagent comprises an antibody molecule with binding specificity for the analyte of interest and the antibody molecule is attached at CH3 domain to the other of the 5’ and 3’ terminus of the first oligonucleotide.

6. The diagnostic device according to claim 2, wherein the first oligonucleotide molecule comprises an aptamer or biomimetic thereof, optionally wherein the first oligonucleotide molecule comprises an aptamer.

7. The diagnostic device according to any one of the preceding claims, wherein the microporous phase material is nitrocellulose.

8. The diagnostic device according to any one of the preceding claims, wherein the covalent binding moiety comprises a N-hydroxysuccinimide ester, aryl azide, phenyl azide, diazirine, psoralen group or anthraquinone.

9. The diagnostic device according to claim 8, wherein the covalent binding moiety comprises anthraquinone C2-dT, or wherein the covalent binding moiety comprises a 6-(4'-azido-2'-nitrophenylamino)hexanoate) group.

10. The diagnostic device according to any one of the preceding claims, wherein the covalent binding moiety is covalently bound to said microporous phase material via photoactivation of a photoreactive functional group.

11. The diagnostic device according to any one of the preceding claims, wherein the long wave electromagnetic radiation (EMR) activable functional group is activated by a wavelength greater than 270nm.

12. The diagnostic device according to claim 11 , wherein the long wave electromagnetic radiation (EMR) activable functional group is activated by a wavelength in the UVA or UVB range.

13. The diagnostic device according to any one of the preceding claims, wherein the covalent binding moiety comprises a photoactivable anthraquinone moiety or a photoactivable nitrophenyl azide.

14. The diagnostic device according to any one of the preceding claims wherein the diagnostic device comprises a detection reagent which is capable of binding the analyte of interest.

15. The diagnostic device according to claim 14 wherein the diagnostic device comprises the detection reagent comprising the second linker moiety, wherein the second linker moiety comprises a second oligonucleotide moiety, wherein said detection reagent is capable of binding the analyte of interest.

16. The diagnostic device according to claim 15, wherein the detection reagent comprises an antibody molecule with binding specificity for the analyte of interest and .wherein the second oligonucleotide molecule attaches to a CH3 domain of the antibody molecule.

17. The diagnostic device according to claim 16, wherein the second oligonucleotide molecule is attached to a CH3 domain of the antibody molecule at either the 5’ or the 3’ end of the second oligonucleotide molecule and is attached to a detectable particle, such as a gold nanoparticle, at the other of the 5’ or 3’ end of the second oligonucleotide molecule.

18. The diagnostic device according to claim 15, wherein the second oligonucleotide has binding specificity for the analyte of interest.

19. The diagnostic device according to claim 15, wherein the second oligonucleotide comprises a detectable particle, such as a gold nanoparticle.

20. The diagnostic device according to claim15, 18 or claim 19, wherein the second oligonucleotide molecule comprises an aptamer or biomimeticthereof, optionally wherein the first oligonucleotide molecule comprises an aptamer.

21. The diagnostic device according to any one of claims 14 to 20, wherein (i) either the capture reagent or the detection reagent comprises an antibody molecule with binding specificity for the analyte of interest, and (ii) the other of the capture reagent and the detection reagent comprises an aptamer with binding specificity for the analyte of interest.

22. The diagnostic device according to claim 21 , wherein the capture reagent comprises an antibody molecule with binding specificity for the analyte of interest and the detection reagent comprises an aptamer with binding specificity for the analyte of interest.

23. The diagnostic device according to claim 21 , wherein the detection reagent comprises an antibody molecule with binding specificity for the analyte of interest and the capture reagent comprises an aptamer with binding specificity for the analyte of interest24. The diagnostic device according to any one of the preceding claims wherein the capture reagent is deposited on the capture zone in a spot of area equivalent to a round reaction site of diameter 100-500 pm.

25. The diagnostic device of claim 24, wherein the spot comprises capture reagent at a concentration in the range 140 pg / pm2to 1400 pg / pm2of the spot.

26. The diagnostic device of claim 25 wherein the capture reagent comprises an antibody molecule.

27. The diagnostic device of claim 24, wherein the capture reagent comprises an aptamer, wherein the spot comprises capture reagent at a concentration in the range 0.140 pg / pm2to 140 pg / pm2of the spot.

28. The diagnostic device according to any one of the preceding claims wherein the diagnostic device comprises a plurality of said capture zones.

29. The diagnostic device according to claim 28 wherein the capture reagent bound to each of the capture zones differs from the capture reagent bound to each of the other capture zones.

30. The diagnostic device according to claim 28 or claim 29, wherein the capture reagents in each capture zone have binding specificity for a different analyte of interest from the capture reagents in one or more of the other capture zones.

31. The diagnostic device according to claim 28 or claim 29, wherein the capture reagents in each capture zone have binding specificity for a different analyte of interest from the capture reagents in each of the other capture zones.

32. The diagnostic device according to any one of claims 28 to 30, wherein the capture reagents in at least a first capture zone have binding specificity for a nucleic acid analyte and the capture reagents in at least a second capture zone have binding specificity for a protein analyte.

33. The diagnostic device according to claim 32, wherein the capture reagents in the first capture zone bind to said nucleic acid analyte via nucleic acid hybridisation and the capture reagents in the second capture zone are aptamers and bind to said protein analyte via aptamer protein interaction.

34. The diagnostic device according to any one of the preceding claims wherein said diagnostic device is a lateral flow device.

35. The diagnostic device according to any one of the preceding claims wherein the diagnostic device includes a calibration zone comprising predetermined different concentrations of calibration molecules for at least one analyte of interest, which in coming into contact with a sample buffer,enable comparison with and quantification of the concentration of the analyte of interest in the capture zone.

36. A diagnostic device comprising: a microporous phase material, a capture reagent bound to a capture zone of the microporous material, wherein the capture reagent is for binding an analyte of interest in a test sample, a calibration zone comprising a plurality of different reference concentrations of calibration molecules for the analyte of interest, wherein, in use, the concentration of the test sample analyte of interest bound to the capture reagent in the capture zone may be determined by comparison of the signal produced by binding of the test sample analyte of interest in the capture zone with the signal produced by one or more of the calibration molecule concentrations in the calibration zone.

37. The diagnostic device according to claim 35 or claim 36, wherein the calibration molecules comprise predetermined concentrations of the analyte of interest bound to the microporous phase material in the calibration zone.

38. The diagnostic device according to claim 35 or claim 36, wherein the calibration molecules comprise predetermined concentrations of said capture reagent known to bind a particular concentration of analyte, said calibration molecules being bound to the microporous phase material in the calibration zone.

39. The diagnostic device according to any one of claims 35-39, wherein the signals produced at a plurality of calibration molecules in the calibration zone can be used to produce a calibration curve against which the signal produced by the analyte in the test sample binding to the capture reagent in the capture zone may be compared, optionally enabling the concentration of analyte in the test sample to be determined.

40. The diagnostic device of claim 36, wherein the device is a device according to any one of claims 1-35.

41. A method of determining the concentration of an analyte of interest in a test sample, said method comprising(i) providing a diagnostic device according to any one of claims 35-40;(ii) bringing the test sample into contact with capture reagent in the capture zone; and(iii) comparing the signal produced by binding of the test sample analyte of interest in the capture zone with the signal produced by one or more of the reference calibration molecule concentrations in the calibration zone.

42. The method according to claim 41 , wherein step (iii) comprises: producing a calibration curve from the signals of calibration molecules in the calibration zone; and comparing the signal produced by the analyte bound to the capture reagent in the capture zone with the calibration curve, thereby enabling the concentration of analyte in the test sample to be determined.

43. A method of manufacturing a diagnostic device, wherein the method comprises:(a) providing a microporous substrate,(b) bringing a capture reagent into contact with said microporous substrate, wherein said capture reagent comprises a linker moiety and a covalent binding moiety at a terminus of the linker moiety, wherein the covalent binding moiety comprises an electromagnetic radiation (EMR)- activable functional group, wherein each linker moiety comprises, or consists of, either an oligonucleotide moiety, a hydrocarbon chain, or an oligonucleotide moiety linked to a hydrocarbon chain, and(c) exposing said microporous substrate to a source of electromagnetic radiation, wherein said exposure causes said capture reagent to covalently bind to said capture zone of said microporous substrate.

44. The method according to claim 43, wherein, during the step of exposure to said electromagnetic radiation, the regions of the microporous substrate other than the predetermined capture zone are masked to prevent EMR- activable functional groups in such zones other than the capture zone being activated.

45. The method according to claim 43 or claim 44, wherein the electromagnetic radiation is visible light, ultraviolet (UV) light, or electron beam irradiation.

46. The method according to any one of claims 43 to 45 wherein the diagnostic device is the diagnostic device of any one of claims 1 to 39.

47. The method according to any one of claims 43 to 46 further comprising a step of washing non-covalently bound capture reagent from the microporous substrate.

48. The method according to any one of claims 43-47 wherein the diagnostic device is a multiplex diagnostic device having a plurality of capture zones, wherein the capture reagents in at least a first of said capture zones have binding specificity for a different analyte of interest from the capture reagents in at least a second of said capture zones.

49. The method according to claim 48, wherein the capture reagents in at least a first capture zone have binding specificity for a nucleic acid analyte and the capture reagents in at least a second capture zone have binding specificity for a protein analyte, optionally wherein the capture reagents in the first capture zone bind to said nucleic acid analyte via nucleic acid hybridisation and the capture reagents in the second capture zone are aptamers and bind to said protein analyte via aptamer protein interaction.

50. The method according to claim 48 or claim 49 wherein the capture reagent bound to each of the capture zones differs from the capture reagent bound to each of the other capture zones.

51. The method according to any one of claims 48-50 wherein the method comprises:(a) bringing a first capture reagent into contact with said microporous substrate, wherein said first capture reagent comprises an oligonucleotidemoiety and a covalent binding moiety at the 5’ or 3’ terminus of the oligonucleotide moiety, wherein the covalent binding moiety comprises an electromagnetic radiation (EMR)-activable functional group,(b) applying a first mask to said microporous substrate, said first mask having a grating corresponding to a first capture zone of said microporous substrate(c) exposing said first capture zone to a source of electromagnetic radiation, wherein the mask prevents exposure of the microporous substrate to said electromagnetic radiation in regions of the microporous substrate other than the first capture zone, wherein said exposure causes said first capture reagent to covalently bind to said first capture zone of said microporous substrate via said EMR-activable functional group,(d) removing said first mask and optionally washing unbound first capture reagent from the microporous substrate;(e) bringing a second capture reagent into contact with said microporous substrate, wherein said second capture reagent comprises a EMR- activable functional group,(f) applying a second mask to said microporous substrate, said second mask having a grating corresponding to a second capture zone of said microporous substrate(g) exposing said second capture zone to a source of electromagnetic radiation, wherein the mask prevents exposure of the microporous substrate to said electromagnetic radiation in regions of the microporous substrate other than the second capture zone, wherein said exposure causes said second capture reagent to covalently bind to said second capture zone of said microporous substrate via said EMR-activable functional group of said second capture reagent ,(h) removing second mask and optionally washing unbound second capture reagent from the microporous substrate; and(i) optionally repeating steps (e) - (h) for third and subsequent capture reagents at third and subsequent capture zones using specific masks for each capture zone.

52. The method according to claim 51 , wherein each of said capture reagents comprises an oligonucleotide moiety specific for a corresponding analyte of interest.

53. The method according to any one of claims 43-51 wherein said diagnostic device is a lateral flow device.

54. The method according to any one of claims 43-52 wherein the diagnostic device comprises one or more of an absorbent pad downstream of said capture zone, a control zone downstream of said capture zone and a quenching zone upstream of said capture zone.

55. The method according to any one of claims 43-54 wherein at least one capture zone comprises a reagent with specificity for an analyte of interest indicative of sepsis Ebola virus or coronavirus.

56. The method according to any one of claims 43-55 wherein reaction of the reagent with an analyte of interest results in a detectable colour change.

57. The method according to any one of claims 43-56, wherein the diagnostic device includes a calibration zone comprising predetermined different concentrations of at least one calibration molecule for the analyte of interest, which in coming into contact with a sample buffer, enable comparison with and quantification of the concentration of the analyte of interest in the capture zone.

58. The method according to claim 57, wherein the calibration molecules comprise predetermined concentrations of the analyte of interest bound to the microporous phase material in the calibration zone.

59. The method according to claim 57, wherein the calibration molecules comprise predetermined concentrations of said capture reagent known to bind a particular concentration of analyte, said calibration molecules being bound to the microporous phase material in the calibration zone.

60. The method according to any one of claims 57-59, wherein the signals produced at a plurality of calibration molecules in the calibration zone may be used to produce a calibration curve against which the signal produced by the analyte in the test sample binding to the capture reagent in the capture zone may be compared, thus enabling the concentration of analyte in the test sample to be determined.

61. A diagnostic device produced using the method of any one of claims 43- 60.

62. A method of detecting the presence of an analyte in a sample, said method comprising contacting a sample comprising said analyte with a diagnostic device according to any one of claims 1-40 or claim 61 or a diagnostic device produced according to the method of any one of claims 43-60 and detecting a colour change in the capture zone.