SARS-CoV-2 surrogate virus neutralization assay test kit

Patent metadata

Field	Value
Publication number	US-11112412-B1
Application number	US-202016939405-A
Country	US
Kind code	B1
Filing date	Jul 27, 2020
Priority date	Mar 25, 2020
Publication date	Sep 7, 2021
Grant date	Sep 7, 2021

Abstract

A kit, composition and method for detection of antibodies to severe acute respiratory syndrome related coronavirus (SARSr-CoV), and for diagnosis of SARSr-CoV infection.

First claim

What is claimed is:

1. A kit for detecting neutralizing antibodies to a severe acute respiratory syndrome-related coronavirus (SARSr-COV), comprising:
(i) a SARS-COV-1 or -2 spike protein or an ACE2-binding fragment of a SARS-COV-1 or -2 spike protein,
(ii) a human ACE2 protein or a SARS-CoV-1 or -2 receptor binding domain (RBD) binding fragment of a human ACE2 protein thereof which binds specifically to the spike protein or the fragment thereof of (i), and
a detection entity comprising a horseradish peroxidase, for detecting interaction between the spike protein or the ACE2-binding fragment of (i) and the human ACE2 protein or RBD binding fragment of (ii),
wherein the spike protein or the ACE2-binding fragment of (i) is conjugated to the detection entity; and the human ACE2 protein or the RBD binding fragment of (ii) is immobilised on a solid support.

2. The kit according to claim 1 ,
wherein the kit is for detecting neutralizing antibodies to SARSr-COV in a sample and the sample is a blood sample, a lymph sample, a saliva sample, or a synovial fluid sample and the SARSr-CoV is SARS-CoV-2.

3. The kit according to claim 1 , wherein the SARSr-CoV is SARS-CoV-2.

4. The kit according to claim 1 , wherein the spike protein or the ACE2-binding fragment of (i) comprises an S1 subunit that comprises an amino acid sequence comprising SEQ ID NO: 12 or 27.

5. The kit according to claim 1 , wherein the kit is for performing a method of analysing a sample for the presence of neutralizing antibodies to a SARSr-CoV, comprising: contacting the sample with (i) the spike protein or the ACE2-binding fragment, and (ii) the human ACE2 protein or the RBD binding fragment, and determining the level of interaction between the spike protein or ACE2-binding fragment of (i) and the human ACE2 protein or the RBD binding fragment of (ii), and
wherein
the spike protein or the ACE2-binding fragment of (i) is at a quantity that is:
(a) sufficient to produce a detectable signal of interaction between the spike protein or the ACE2-binding fragment of (i) and the human ACE2 protein or the RBD binding fragment of (ii) in the absence of the sample, and/or
(b) less than or equal to, in molar ratio, the quantity of the antibodies to the SARSr-CoV in the sample, and/or
(c) a minimal quantity required to produce a detectable signal of interaction between the spike protein or the ACE2-binding fragment of (i) and the ACE2 protein or the RBD binding fragment of (ii) in the absence of the sample.

6. The kit according to claim 1 , wherein the RBD comprises an amino acid sequence comprising SEQ ID NO: 13 or 26.

7. The kit according to claim 1 , wherein the ACE2 protein or the RBD-binding fragment of (ii) comprises an ACE2 protein extracellular domain having an amino acid sequence comprising SEQ ID NO: 17 or 30.

8. The kit according to claim 5 , wherein the spike protein or the ACE2-binding fragment of (i) comprises an S1 subunit that comprises an amino acid sequence comprising SEQ ID NO: 12 or 27.

9. The kit according to claim 1 , wherein the kit is for performing a method of analysing a sample for the presence of neutralizing antibodies to a SARSr-CoV, comprising: contacting the sample with (i) the spike protein or the ACE2-binding fragment, and (ii) the human ACE2 protein or the RBD binding fragment, and determining the level of interaction between the spike protein or ACE2-binding fragment of (i) and the human ACE2 protein or the RBD binding fragment of (ii).

10. The kit according to claim 9 , wherein the sample is a blood sample, a lymph sample, a saliva sample, or a synovial fluid sample.

11. The kit according to claim 9 , wherein the SARSr-CoV is SARS-CoV-2.

12. The kit according to claim 5 , wherein the RBD comprises an amino acid sequence comprising SEQ ID NO: 13 or 26.

13. The kit according to claim 5 , wherein the ACE2 protein or the RBD-binding fragment of (ii) comprises an ACE2 protein extracellular domain having an amino acid sequence comprising SEQ ID NO: 17 or 30.

14. The kit according to claim 9 , wherein the spike protein or the ACE2-binding fragment of (i) comprises an S1 subunit that comprises an amino acid sequence comprising SEQ ID NO: 12 or 27.

15. The kit according to claim 5 , wherein the sample is a blood sample, a lymph sample, a saliva sample, or a synovial fluid sample.

16. The kit according to claim 9 , wherein the RBD comprises an amino acid sequence comprising SEQ ID NO: 13 or 26.

17. The kit according to claim 9 , wherein the ACE2 protein or the RBD binding fragment of (ii) comprises an ACE2 protein extracellular domain having an amino acid sequence comprising SEQ ID NO: 17 or 30.

18. The kit according to claim 9 , wherein the spike protein or the ACE2-binding fragment of (i) is at a quantity that is (a) sufficient to produce a detectable signal of interaction between the spike protein or the ACE2-binding fragment (i) and the ACE2 protein or the RBD binding fragment of (ii) in the absence of the sample, and (b) less than or equal to, in molar ratio, the quantity of the antibodies to the SARSr-CoV in the sample.

19. The kit according to claim 9 , wherein the spike protein or the ACE2-binding fragment of (i) is at a minimal quantity required to produce a detectable signal of interaction between the spike protein or the ACE2-binding fragment of (i) and the ACE2 protein or the RBD binding fragment of (ii) in the absence of the sample.

20. The kit according to claim 5 , wherein the spike protein or the ACE2-binding fragment of (i) is at a quantity that is: (a) sufficient to produce a detectable signal of interaction between the spike protein or the ACE2-binding fragment of (i) and the ACE2 protein or the RBD binding fragment of (ii) in the absence of the sample.

21. The kit according to claim 5 , wherein the spike protein or the ACE2-binding fragment of (i) is at a quantity that is: (c) a minimal quantity required to produce a detectable signal of interaction between the spike protein or the ACE2-binding fragment of (i) and the ACE2 protein or the RBD binding fragment of (ii) in the absence of the sample.

22. The kit according to claim 20 , wherein the sample is a blood sample, a lymph sample, a saliva sample, or a synovial fluid sample and the SARSr-CoV is SARS-CoV-2.

23. The kit according to claim 21 , wherein the sample is a blood sample, a lymph sample, a saliva sample, or a synovial fluid sample and the SARSr-CoV is SARS-CoV-2.

24. The kit according to claim 22 , wherein the spike protein or the ACE2-binding fragment of (i) comprises an S1 subunit that comprises the amino acid sequence set forth in SEQ ID NO: 12 or 27, or the RBD comprises an amino acid sequence comprising SEQ ID NO: 13 or 26, or the ACE2 protein or the RBD binding fragment of (ii) comprises an ACE2 protein extracellular domain having an amino acid sequence comprising SEQ ID NO: 17 or 30.

25. The kit according to claim 23 , wherein the spike protein or the ACE2-binding fragment of (i) comprises an S1 subunit that comprises the amino acid sequence set forth in SEQ ID NO: 12 or 27, or the RBD comprises an amino acid sequence comprising SEQ ID NO: 13 or 26, or the ACE2 protein or the RBD binding fragment of (ii) comprises an ACE2 protein extracellular domain having an amino acid sequence comprising SEQ ID NO: 17 or 30.

26. The kit according to claim 9 , wherein the sample is a blood sample, a lymph sample, a saliva sample, or a synovial fluid sam

Assignees

• Nat Univ Singapore

Inventors

• Wang Linfa

Classifications

• C07K16/104

  Severe acute respiratory syndrome coronavirus 2 [SARS‐CoV‐2] · CPC title

• C07K16/102

  Coronaviridae (F) · CPC title

• A61J1/00

  Containers specially adapted for medical or pharmaceutical purposes (capsules or the like for oral use A61J3/07; specially adapted for surgical or diagnostic appliances or instruments A61B50/30; containers for radioactive substances G21F5/00) · CPC title

• G01N33/6854 Primary

  Immunoglobulins · CPC title

• G01N33/56983 Primary

  Viruses · CPC title