METHODS OF TAGGING PARTICLES FOR MULTIPLEXED FUNCTIONAL SCREENING
The present invention relates generally to the fields of cell biology and laboratory diagnostics, and particularly to general compositions and of uniquely tagged particles linked to moieties of known properties and methods of making tagged, functionalized particles. Additionally, the invention relates to methods of screening a collection of tagged functionalized particles.
1 . A method of screening a collection of uniquely tagged, functionalized particles to identify a functional moiety or combination of functional moieties having a desired property or properties, the method comprising:
preparing a collection of tagged, functionalized particles comprising one or more functional moieties and a unique tag linked to the particle;
introducing the collection of tagged, functionalized particles into an assay to select for a specific property or properties;
isolating the tagged, functionalized particles that manifest the desired property or properties;
identifying the tags of the isolated tagged functionalized particles; and
determining the functional moiety or combination of functional moieties from the identity of the tag.
2 . The method of claim 1 , wherein the assay is in vitro or in vivo.
3 . The method of claim 1 , wherein the desired property or properties of the functional moiety or combination of functional moieties comprise a location, a concentration, a binding affinity, a pharmacokinetic property, a pharmacodynamic property, or a chemical property.
4 . The method of claim 1 , wherein
if the tags are nucleic acids, identifying the tags comprises nucleotide sequencing;
if the tags are peptides or mass-encoded tags, identifying the tags comprises mass spectrometry; and
if the tags are fluorescent dyes, quantum dots or nanodiamonds, identifying the tags comprises flow cytometry or microscopy.
5 . The method of claim 1 , wherein the particle is a liposome.
6 . The method of claim 5 , wherein the one or more functional moieties is covalently linked to the liposome.
7 . The method of claim 5 , wherein the one or more functional moieties is non-covalently embedded within the liposome.
8 . The method of claim 5 , wherein the unique tag is covalently linked to the liposome.
9 . The method of claim 5 , wherein the unique tag is non-covalently embedded within the liposome.
10 . The method of claim 1 , wherein the unique tag is a nucleic acid, a peptide, an optically-encoded tag or a mass-encoded tag.
11 . The method of claim 1 , wherein the unique tag is a nucleic acid comprising DNA, RNA or XNA.
12 . The method of claim 11 , wherein the nucleic acid comprises 5-150 nucleotides of known sequence.
13 . The method of claim 11 , wherein the nucleic acid is single stranded.
14 . The method of claim 11 , wherein the nucleic acid is double stranded.
15 . The method of claim 1 , wherein the unique tag is a peptide comprising a short peptide, a peptoid, or a specific pattern of isotope labeled peptide.
16 . The method of claim 15 , wherein the peptide or peptoid comprises 5-20 amino acids of known sequence.
17 . The method of claim 1 , wherein the unique tag is an optically-encoded tag comprising a fluorescent dye, quantum dot, polymer dot, nanodiamond, FRET system or combination thereof.
18 . The method of claim 1 , wherein the unique tag is a mass-encoded tag comprising one or more lanthanide atom embedded within or attached to the surface of the particle.
19 . The method of claim 1 , wherein the at least one functional moiety is a monoclonal antibody, a polyclonal antibody, single chain antibody, a single domain antibody, a bi-specific antibody, an affibody molecule, a peptide, a peptoid, an aptamer, a small molecule or a chemical compound.
20 . The method of claim 1 , wherein the functionalized particles comprise a first functional moiety and a second functional moiety, wherein the second functional moiety is a monoclonal antibody, a polyclonal antibody, single chain antibody, a bi-specific antibody, a small molecule or a chemical compound, and the first functional moiety is different than the second functional moiety.