Method of isolating analytes from a sample
The current invention is a capture-particle comprising: a) a molecular sieve portion; and b) an analyte binding portion; wherein the molecular sieve portion, analyte binding portion or both further comprise a cross-linked region having modified porosity. Capture particles wherein the molecular sieve portion, analyte binding portion or both comprise pore dimensions sufficient to exclude molecules larger than about 60 kDa. These particles are useful in purification and diagnostic methods. Kits comprising the capture particles are also described.
1. A capture-particle consisting of:
a) an outer molecular sieve portion; and
b) an inner analyte binding portion with reactive organic affinity molecules configured for binding and sequestering analytes such that there are no dyes used for signaling detection;
wherein the molecular sieve portion, analyte binding portion or both further comprise a cross-linked region having porosity physically configured to permit passage of analytes of a defined molecular mass or weight such that there is no chemical cleaving.
2. The capture-particle of claim 1 wherein said analyte binding portion comprises at least one type of moiety capable of chemically or electrostatically binding or sequestering an analyte.
3. The capture-particle of claim 1 wherein the analyte binding portion comprises a carboxy group, amine group, lipid, phosphoprotein, phospholipids, amide group, hydroxyl group, ester group, acrylic group, thiol group, acrylic acid, antibodies, binding proteins, binding pairs, metals, chelating agents, nucleic acids, aptamers, enzyme-binding pockets, lectins, pharmacologic agent, synthetic peptides, antibody fragments, hydrophobic surface, hydrophyllic surface, any derivatives thereof or a combination thereof.
4. The capture-particle of claim 1 further comprising an analyte bound to the analyte binding portion, said analyte comprising: organic molecules, inorganic molecules, polypeptides, carbohydrates, nucleic acids, lipids, derivatives thereof or any combination thereof.
5. The capture-particle of claim 1 wherein the molecular sieve portion is an outer shell enclosing an inner core, said inner core comprising the analyte binding portion.
6. The capture-particle of claim 1 having an average particle size radius of less than about 100μ.
7. The capture-particle of claim 1 wherein the molecular sieve portion, analyte binding portion or both comprise: polyacrylamide, poly(Nisopropylacrylamide), N-alkyl substituted polyacrylamide, poly(N-vinylalkylamide), poly(methacrylic acid), poly(benzyl glutamate), ply(2-ethylacrylic acid), poly(4-vinylpyridine), derivatives thereof or any combination thereof.
8. The capture-particle of claim 1 wherein the cross-linked region comprises N,N′-methylenebisacrylamide, ethylene glycol dimethacrylate, tetraethylene glycol dimethacrylate, poly(ethyleneglycol)dimethacrylate or any combination thereof.
9. The capture-particle of claim 1 wherein the molecular sieve portion, analyte binder portion or both comprise a hydrogel.
10. The capture-particle of claim 1 wherein the molecular sieve portion, analyte binding portion or both have an average pore size of between about 2 to about 20 nm.
11. The capture-particle of claim 1 the molecular sieve portion, analyte binding portion or both have an average pore size of less than about 100 nm.
12. The capture-particle of claim 1 wherein said capture-particle releases a bonded or sequestered analyte, changes volume or both when exposed to a physical or chemical treatment.
13. The capture-particle of claim 12 wherein the physical or chemical treatment comprises exposure to: electrical charge, hydrostatic pressure, change in pH, change in temperature, acidic agent, basic agent, UV, ultrasound, x-ray, or a combination thereof.
14. The capture-particle of claim 1 having the ability to uptake a 1404 Da peptide with substantially no uptake of albumin.
15. The capture-particle of claim 1 having the ability to uptake insulin.
16. A kit for the diagnosis, prognosis or monitoring of a disease state consisting of:
a container for collecting a fluid comprising analytes indicative of said disease state; and
an amount of capture-particles for uptake and removal of said analytes, said capture-particles consisting of:
a) an outer molecular sieve portion; and
b) an inner analyte binding portion with reactive organic affinity molecules configured for binding and sequestering analytes such that there are no dyes used for signaling detection;
wherein the molecular sieve portion, analyte binding portion or both further comprise a cross-linked region having porosity physically configured to permit passage of analytes of a defined molecular mass or weight such that there is no chemical cleaving.