Layered particles for retrieving DNA released from cells from a gastrointestinal tract sample and methods for making and using them
View Patent ↗In alternative embodiments, the invention provides compositions such as devices, pills, beads, capsules, products of manufacture, particles, microparticles, nanoparticles, gels, liquid gels, liquid gel capsules, capsules, tablets, geltabs, liquids, sprays, emulsions, suspensions, pastes or yogurts, for the detection and isolation of biomarkers, nucleic acids, proteins or peptides, proteoglycans, lipids, fats, sugars or polysaccharides in the gastrointestinal tract for e.g., detecting the presence of particular exogenous or endogenous nucleic acids, e.g., DNA or RNA, or proteins, in the gastrointestinal tract, for example, to diagnose the presence of an infectious or exogenous agent such as a virus, a fungus, a parasite, a bacteria, intestinal helminths and protozoan parasites, and the like, or a biomarker such as a cancer-causing or cancer-predisposing allele, e.g., mutations of the KRAS2 oncogene in pancreatic cancer. In alternative embodiments, compositions of the invention comprise magnetic particles such as a magnetically-responsive microparticle or nanoparticle; a superparamagnetic bead or polystyrene bead; a superparamagnetic fine particle; a ferrimagnetic particle; or, a magnetic microsphere, nanosphere, microbead or nanobeads. In alternative embodiments, the invention provides methods for detecting, retrieving, capturing or isolating a sample of a nucleic acid, or an anionic, cationic or hydrophobic protein or peptide, a mucin, a phosphoprotein, a proteoglycan or a polysaccharide, in vivo.
1. A layered particle for retrieving DNA released from cells from a human gastrointestinal sample comprising:
an outer shell or layer and an inner core or layer,
wherein the outer shell or layer comprises an outer surface that is hydrophilic, and is neutral in charge, and
the outer shell or layer comprises pores having a pore size of between 1 nm and 100 nm;
and the inner core or layer comprises positively charged cationic polymers or molecules attached onto its outer surface, wherein the positively charged cationic polymers or molecules comprise
a positively charged branched, cross-linked dextran or a quaternized positively charged branched, cross-linked polyethylenimine, or a combination thereof.
2. The layered particle of claim 1 , wherein the inner core or layer further comprises a quaternary, a tertiary, a primary or a secondary amine, or a combination thereof, or a guanidine, a quaternized polyethylenimine (PEI), a pentamethylguanidine, a heptamethylisobiguanide or a Hunig's base or a mixture thereof.
3. The layered particle of claim 1 , wherein the outer shell or layer and/or an inner core or layer further comprises a plurality of magnetic molecules or particles.
4. The layered particle of claim 1 ,
wherein the positively charged cationic inner core or layer comprises a nanoparticle, microparticle or polymer that is positively charged (cationic).
5. The layered particle of claim 1 , formulated for ingestion for a human.
6. A gel, a capsule, a tablet, a liquid, a spray, an emulsion, a suspension, a paste, a yogurt, or a geltab, comprising: a layered particle of claim 1 .
7. The layered particle of claim 1 , wherein the pores have a pore size of 1.5 nm.
8. The layered particle of claim 5 , wherein the layered particle is encapsulated in:
(a) an enteric coating, wherein optionally the enteric coating comprises a coating resistant in the gastric pH but removable in human duodenal pH, or dissolves or disintegrates in human duodenal pH; or
(b) a capsule, tablet or geltab that dissolves or disintegrates in human duodenal pH, or at a pH of between pH 5 and pH 6.
9. The layered particle of claim 3 , wherein the plurality of magnetic molecules or particles comprise a magnetically-responsive microparticle or nanoparticle; a superparamagnetic bead or polystyrene bead; a superparamagnetic fine particle; a ferrimagnetic particle; a magnetic microsphere; a magnetic microbead; or, a magnetic nanobead.
10. The gel, capsule, tablet, liquid, spray, emulsion, suspension, paste, yogurt, or geltab of claim 6 , further comprising a DNase inhibitor, an EGTA, an EDTA, an actin, a polycations, a flavoring agent or a coloring agent, or a mixture thereof or a combination thereof.
11. The layered particle of claim 1 , wherein the pores have a pore size of 2 nm, 2.5 nm, 3 nm, 3.5 nm or 4 nm.
12. The layered particle of claim 11 , wherein the pores have a pore size of 5 nm, 10 nm or 15 nm.
13. The layered particle of claim 12 , wherein the pores have a pore size of 20 nm, 25 nm, 30 nm or 40 nm.
14. The layered particle of claim 13 , wherein the pores have a pore size of 50 nm, 60 nm, 70 nm, 80 nm, or 90 nm.
15. The layered particle of claim 1 , wherein the positively-charged branched, cross-linked dextran or the positively-charged branched, cross-linked quaternized polyethylenimine (PEI) comprises positively-charged branched, cross-linked dextran or positively-charged branched, cross-linked quaternized polyethylenimine (PEI) of between 10 to 500 kDa.
16. The layered particle of claim 1 , wherein the inner core or layer has attached onto its outer surface molecules comprising a positively charged branched, cross-linked dextran.
17. The layered particle of claim 1 , wherein the inner core or layer has attached onto its outer surface molecules comprising a positively charged branched, cross-linked quaternized polyethylenimine (PEI).
18. The layered particle of claim 2 , wherein the inner core or layer further comprises a quaternary, a tertiary, a primary or a secondary amine, or a combination thereof.
19. The layered particle of claim 2 , wherein the inner core or layer further comprises a guanidine, a quaternized polyethylenimine (PEI), a pentamethylguanidine, a heptamethylisobiguanide or a Hunig's base or a mixture thereof.