METHODS. PARTICLES, AND ASSAY KITS FOR IDENTIFYING PRESENCE OF BIOLOGICAL PARAMETERS

20130123145

13/726021

December 22, 2012

Multiplexed assays are disclosed for detection of duster differentiation 4 (CD4) glycoprotein expressed on the cell surface of I-helper cells, monocytes, macrophages, and dendritic cells; cluster differentiation 25 (CD25), a type transmembrane protein present on activated T-cells, activated B-cells, thymocytes, myeloid precursors, and oligodendrocytes; and Forkhead box P3 (FOXP3), an intracellular protein involved in immune system responses in cell cultures, tissues samples, humans, and biological samples. The multiplexed assays can be used to detect 1r-cells, activated I-cells, and other similar cell types (e.g. natural T regulatory cells, adaptive/induced T regulatory T cells). The multiplexed assays employ quantum dots of various shapes, types, compositions, coatings, sizes, ligands and other such characteristics.

NNANOAXIS, LLC (Clarence, NY, US)

1. An assay kit comprising: a set of hydrophilic coated nanoparticles paired to K biological materials that respectively form L nanoplexes that respectively bind to M predefined targeting moieties that correspond to N biological parameters and luminesce at respective wavelengths that correspond to the respective predefined biological parameter to determine P predefined target cell types, wherein K, L, M, N, and P are integers.

2. The assay kit of claim 1, wherein the hydrophobic coated nanoparticles are at least one of a: nanocrystal, quantum dot, spherical nanocrystal, non-spherical nanocrystal, spherical quantum dot, non-spherical quantum dot, tetrapod quantum dot, multi-leg luminescent nanomaterial, doped nanoparticle, polymer encapsulated quantum dot, or nanoparticle that exhibit luminescent properties.

3. The assay kit of claim 1, wherein the quantum dot is at least one of a: heavy metal-free quantum dot, cadmium-free quantum dot, phosphorous quantum dot, or biocompatible quantum dot.

4. The assay kit of claim 1, wherein the hydrophilic coat is any one or more of: Pluronic F127, silicon, micelle, glass, polymeric oxide, oxide of phosphorous, or polymeric ligands, amphiphilic ligand, or hydrophilic thiol compound.

5. The assay kit of claim 1 wherein the predefined targeting moities are at least one of an: antibody, monoclonal antibody, polyclonal antibody, nucleic acid, monomeric nucleic acid, oligomeric nucleic acid, protein, polysaccharide, sugar, peptide, drug, RNA, DNA, microRNA, or plasmid.

6. The assay kit of claim 1, wherein the hydrophilic coated nanoparticles are produced in a continuous flow chemistry process.

7. The assay kit of claim 1, wherein the hydrophilic coated nanoparticles are non-spherical quantum dots comprising a core material surrounded by a shell material that is produced using a continuous flow chemistry process.

8. The assay kit of claim 1, wherein the hydrophilic coated nanoparticles are non-spherical quantum dots comprising a core material surrounded by a shell material that is aqueous solubilized and produced using a continuous flow chemistry process.

9. The assay kit in claim 1, wherein the hydrophilic coated nanoparticles are non-spherical quantum dots comprising a core material surrounded by a shell material that is aqueous solubilized, paired to an antibody, and produced a using continuous flow chemistry process

10. The assay kit of claim 1, wherein the biological material is at least one of of: CD4 antibody, CD25 antibody, CD17 anitbody, TLR antibody, FoxP3 antibody, or methyl antibody.

11. The assay kit of claim 1, wherein the predetermined target cell types are any one or more of: T regulatory cells, TH17 cells, T-cells or Toll-like receptors present on cells.

12. The assay kit of claim 1, further comprising at least two nanoplex, wherein each nanoplex emits at a different wavelength.

13. The assay kit of claim 1, wherein the nanoplexes are packaged as a Z-plex assay kit, wherein Z is an integer.

14. The assay kit of claim 1, adapted for use in at least one of a lateral flow assay, ELISA, sandwich ELISA, microarray, piezoarray, Western blot, flow cytometry, or UV spectromphotometer.

15. A method of producing an assay kit comprising: coating a set of nanoparticles to form a set of hydrophilic coated nanoparticles; pairing the V set of hydrophilic nanoparticles of one or more wavelengths to W biological materials to form X nanoplexes; V, W and X are integers.

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