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US 11,054,429 B1Granted
SARS-CoV-2 surrogate virus neutralization test based on antibody-mediated blockage of ACE2-spike protein binding
AssigneeNational University of Singapore
InventorLinfa Wang
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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.
Claims26
A method of analysing a sample for the presence of antibodies to a SARSr-CoV, comprising:
- contacting the sample with:
- (i) a spike protein of a severe acute respiratory syndrome-related coronavirus (SARSr-CoV) or a S1 subunit of a SARSr-CoV spike protein, wherein the spike protein or the S1 subunit comprises the receptor binding domain (RBD) of the SARSr-CoV spike protein, and (ii) an ACE2 protein or an extracellular domain of an ACE2 protein a fragment thereof or which binds specifically to the spike protein or the S1 subunit of (i), and
- determining the level of interaction between the spike protein or the S1 subunit of (i) and the ACE2 or the ACE2 protein extracellular domain of (ii),
- wherein the spike protein or the ACE2-binding fragment of (i) or ACE2 or the ACE2 protein extracellular domain of (ii) is conjugated to a detection entity, for determining the interaction between (i) and (ii);
- and
- (a) spike protein or the S1 subunit of (i) is conjugated to the detection entity, and wherein the ACE2 or the ACE2 protein extracellular domain of (ii) is immobilised on a solid support;
- or
- (b) ACE2 or the ACE2 protein extracellular domain of (ii) is conjugated to the detection entity, and wherein spike protein or the S1 subunit of (i) is immobilised on a solid support.
5 more elements
A method of detecting in a sample the presence of an antibody that binds to an receptor binding domain (RBD) of a spike protein of a severe acute respiratory syndrome-related coronavirus (SARSr-CoV), the method comprising:
- a) contacting the sample with a spike protein of SARSr-CoV or a subunit or fragment of the spike protein, wherein the spike protein or the subunit or the fragment comprises an RBD, and wherein the spike protein or subunit or fragment is linked to a detection entity, to produce a mixture,
- b) incubating the mixture to allow the spike protein or a subunit or fragment thereof to interact with any antibody which may exist in the sample to bind to the RBD,
- c) contacting the mixture from step b) with a solid support on which is immobilized an ACE2 protein or an extracellular domain or a fragment of the ACE2 protein which binds specifically to the RBD, and
- d) detecting the presence or absence of a signal from the detection entity, wherein the absence of a signal indicates that the sample contains an antibody that binds to the RBD of the SARSr-CoV.
1 more element
Dependencies are inferred from the claim text.