Assay method for the detection of VCAM-1 and alpha-2-macroglobulin in blood
AssigneeProcisedx Inc.
InventorsHongyu Chen, Limin Liu, Stefan Westin, Michael Hale, Larry Mimms
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Abstract
An assay method for detecting the presence or amounts of VCAM-1 and A2M in a sample using fluorescence resonance energy transfer (FRET).
Claims20
A homogeneous assay method for detecting the presence or amount of vascular cell adhesion protein 1 (VCAM-1) using only a sandwich assay and alpha-2-macroglobulin (A2M) using only a competitive assay in a sample, the method comprising:
- in the sandwich assay, contacting the sample with a first anti-VCAM-1 antibody, which binds to a first epitope of VCAM-1, wherein the first anti-VCAM-1 antibody is labeled with a first donor fluorophore;
- contacting the sample with a second anti-VCAM-1 antibody, which binds to a second epitope of VCAM-1, wherein the second anti-VCAM-1 antibody is labeled with a first acceptor fluorophore;
- in the competitive assay, contacting the sample with an anti-A2M antibody, which binds to an epitope of A2M, wherein the anti-A2M antibody is labeled with a second donor fluorophore;
- contacting the sample with an isolated A2M protein, wherein the isolated A2M protein is labeled with a second acceptor fluorophore;
- incubating the sample for a time sufficient to obtain dual antibody labeled VCAM-1 and antibody labeled A2M;
- and
- exciting the sample having dual antibody labeled VCAM-1 and antibody labeled A2M using one or more light sources to detect a single fluorescence emission signal associated with fluorescence resonance energy transfer (FRET) for the first acceptor fluorophore and a single fluorescence emission signal associated with FRET for the second acceptor fluorophore,
- wherein the first and second acceptor fluorophores are different.
A homogeneous assay method for detecting the presence or amount of vascular cell adhesion protein 1 (VCAM-1) using only a sandwich assay and alpha-2-macroglobulin (A2M) using only a competitive assay in a sample, the method comprising:
- in the sandwich assay, contacting the sample with a first anti-VCAM-1 antibody, which binds to a first epitope of VCAM-1, wherein the first anti-VCAM-1 antibody is labeled with a first donor fluorophore;
- contacting the sample with a second anti-VCAM-1 antibody, which binds a second epitope of VCAM-1, wherein the second anti-VCAM-1 antibody is labeled with a first acceptor fluorophore;
- in the competitive assay, contacting the sample with an anti-A2M antibody, which binds an epitope to A2M, wherein the anti-A2M antibody is labeled with a second acceptor fluorophore;
- contacting the sample with an isolated A2M protein, wherein the isolated A2M protein is labeled with a second donor fluorophore;
- incubating the sample for a time sufficient to obtain dual antibody labeled VCAM-1 and antibody labeled A2M;
- and
- exciting the sample having dual antibody labeled VCAM-1 and antibody labeled A2M using one or more light sources to detect a single fluorescence emission signal associated with fluorescence resonance energy transfer (FRET) for the first acceptor fluorophore and a single fluorescence emission signal associated with FRET for the second acceptor fluorophore, wherein the first and second acceptor fluorophores are different.
Dependencies are inferred from the claim text.