Assay methods involving dissociable nanoparticles
Among other things, the present invention provides assay methods for detecting or quantifying one or more analytes in a sample, which involve the use of dissociable nanoparticles that comprise one or more signaling agents (e.g., the nanoparticles conceal or partially conceal the signaling agents).
1. A method of detecting an analyte, comprising:
(i) incubating a sample suspected of having an analyte of interest with a binding agent specific to the analyte under conditions that permit binding between the analyte and the binding agent; wherein the binding agent is associated with a nanoparticle comprising a first signaling agent and a second signaling agent; wherein the first signaling agent is a metalorganic species catalyst comprising a metal-tetraamidomacrocyclic ligand complex, and the second signaling agent is a pre-chemiluminophore;
(ii) dissociating the nanoparticle bound to the analyte, if any, to release the signaling agents such that it results in a signal change; and
(iii) determining presence or quantity of the analyte in the sample based on the signal change.
2. The method of claim 1 , wherein the nanoparticle further comprises one or more matrix-forming agents providing a matrix, wherein the signaling agents are embedded in the matrix.
3. The method of claim 1 , wherein the binding agent is selected from the group consisting of antibodies or antigen-binding fragments thereof, enzymes, oligonucleotides, DNA, RNA, PNA, or LNA, proteins, peptides, polypeptides, receptors, ligands, small molecules, aptamers, polysaccharides, plastibodies, affibodies, camelids, fibronectins, and a combination thereof.
4. The method of claim 1 , wherein the nanoparticle comprises one or more functional groups for conjugating the nanoparticle to the binding agent.
5. The method of claim 4 , wherein the functional group is linked to the outer surface via polyethylene glycol (PEG).
6. The method of claim 1 , wherein step (ii) and step (iii) are performed simultaneously in a solution.