Sample preparation compositions, devices, systems and methods

Patent metadata

Field	Value
Publication number	US-11975306-B2
Application number	US-202017016140-A
Country	US
Kind code	B2
Filing date	Sep 9, 2020
Priority date	Sep 10, 2019
Publication date	May 7, 2024
Grant date	May 7, 2024

Abstract

The present specification relates to compositions, devices, apparatus, methods, kits and systems for sample preparation (e.g., separation, reduction or removal of small molecules from biomolecules in a sample). Exemplary small molecules that can be separated, reduced or removed have a molecular weight range of <2000 Da. and may include, but are not limited to, dyes, biotin, affinity tags, crosslinkers, reducing agents, labels, nanoparticles, radioactive ligands, mass tags, unreacted molecules and combinations, intermediates and derivatives of the foregoing. Exemplary biomolecules present in a sample, include but are not limited to, proteins, glycoproteins, antibodies, peptides, nucleic acids, polysaccharides, carbohydrates and lipids. Methods, compositions, kits, devices, apparatus and systems of the disclosure may advantageously provide superior separation of small molecule contaminants and additionally reduce time and expenses related to separation of small molecules from larger biomolecules in samples. Biomolecules separated as set forth herein are amenable to better downstream processing.

First claim

What is claimed is:

1. A composition for separating one or more small molecules from a sample, the composition comprising:
a first size exclusion support comprising a polyacrylamide, a cellulose material, a hydroxyethyl cellulose, and/or derivatives thereof; and
a first moiety that is immobilized onto the first size exclusion support, wherein the first moiety can associate with the one or more small molecules to separate the one or more small molecules from a sample.

2. The composition of claim 1 , comprising a second moiety that is different from the first moiety, wherein the second moiety is immobilized onto the first size exclusion support.

3. The composition of claim 1 , further comprising a second size exclusion support that is different from the first size exclusion support.

4. The composition of claim 3 , wherein the second size exclusion support comprises a dextran polymer or agarose.

5. The composition of claim 1 , wherein the first moiety comprises a polysaccharide, a dextran, a polyethylene glycol polymer, an amine-containing polymer, a polyaminoacid, a lipopolysaccharide, an antibiotic, a chelating group, a magnetic particle, a paramagnetic particle, a functional group, an ion-exchanger, or any combination thereof.

6. The composition of claim 5 , wherein the amine-containing polymer is a poly(ethylene glycol)diamine, a polyethylenediamine, a polyethyleneimine that is linear, or a polyethyleneimine that is branched.

7. The composition of claim 6 , wherein the polyethyleneimine that is linear, is diethylenediamine.

8. The composition of claim 5 , wherein the first moiety is a dextran.

9. The composition of claim 8 , wherein the dextran has a molecular weight in the range of from about 6 kDa-2800 kDa.

10. The composition of claim 1 , wherein the first moiety associates with the one or more small molecules by charge interaction, hydrophilic interactions, hydrophobic interactions, affinity interaction, hydrogen bonding, or Van der Waals forces.

11. The composition of claim 1 , wherein the one or more small molecules have a molecular weight range of <2000 Da.

12. The composition of claim 1 , wherein the one or more small molecules independently are selected from an unreacted or partially reacted label or nonoparticle, or a derivative thereof; a dye or derivative thereof; a radioactive ligand or an intermediate thereof; a mass tag; a metal; biotin or a derivative thereof; a crosslinker or derivative thereof; a reducing agent or derivative thereof; or any combination thereof.

13. The composition of claim 1 , wherein the sample comprises a biomolecule, a protein a glycoprotein, an antibody, a peptide, a nucleic acid, a polysaccharide, a carbohydrate or a lipid and wherein contacting the sample with the composition substantially reduces a quantity of the one or more small molecules in the sample.

14. A device for separating one or more small molecules from a sample, the device comprising:
a) a container comprising a composition for separating one or more small molecules from a sample, the composition comprising (i) a first size exclusion support comprising a polyacrylamide, a cellulose material, a hydroxyethyl cellulose, and/or derivatives thereof; and (ii) a first moiety that is immobilized onto the first size exclusion support; wherein the first moiety can associate with the one or more small molecules to separate the one or more small molecules from the sample; and
b) a receptacle located below the container.

15. The device of claim 14 , operably configured to be subject to a gravity flow, a centrifugal force, a positive pressure, a negative pressure, vacuum, or any combination thereof.

16. The device of claim 14 , wherein the container is a columnar container, a tube, a multi-well tube, a multi-well plate, or a multi-well filter plate.

17. A system for separating one or more small molecules from a sample, the system comprising:
a) a container comprising a composition for separating one or more small molecules from a sample, the composition comprising (i) a first size exclusion support comprising a polyacrylamide, a cellulose material, a hydroxyethyl cellulose, and/or derivatives thereof; and (ii) a first moiety that is immobilized onto the first size exclusion support, wherein the firtst moiety can associate with the one or more small molecules to separate the one or more samll molecules from the sample;
b) a receptacle located below the container; and
c) a means to subject the container and receptacle to a gravity flow, a centrifugal force, a positive pressure, a negative pressure, vacuum, or any combinations thereof.

18. A kit for separating a biomolecule from one or more small molecules comprising:
the device of claim 14 , wherein the device is operably configured to be subject to a gravity flow, a centrifugal force, a positive pressure, a negative pressure, vacuum, or any combination thereof.

19. A method for separating at least one biomolecule from one or more small molecules, the method comprising:
a) applying a sample comprising the at least one biomolecule to a composition comprising (i) a first size exclusion support comprising a polyacrylamide, a cellulose material, a hydroxyethyl cellulose, and/or derivatives thereof; and (ii) a first moiety that is immobilized onto the first size exclusion support; and
b) subjecting the container to a gravity flow, a centrifugal force, a positive pressure, a negative pressure, a vacuum, or any combination thereof,
wherein the at least one biomolecule in the sample is excluded through the composition and is collected as a flow through, and wherein the one or more small molecules associates with the first one moiety and is thereby separated from the at least one biomolecule.

Assignees

• Pierce Biotechnology Inc

Inventors

• Ganapathy Ramesh
• Etienne Christopher
• Etnyre Deven
• Salunkhe Ashok
• Deshpande Atul

Classifications

• B01J20/3208

  Polymeric carriers, supports or substrates · CPC title

• B01J20/24 Primary

  Naturally occurring macromolecular compounds, e.g. humic acids or their derivatives · CPC title

• B01D15/34 Primary

  Size-selective separation, e.g. size-exclusion chromatography; Gel filtration; Permeation · CPC title

• B01J20/3042

  Use of binding agents; addition of materials ameliorating the mechanical properties of the produced sorbent · CPC title

• C07K1/14

  Extraction; Separation; Purification · CPC title