Sample preparation compositions, devices, systems and methods

Patent metadata

Field	Value
Publication number	US-2021069673-A1
Application number	US-202017016140-A
Country	US
Kind code	A1
Filing date	Sep 9, 2020
Priority date	Sep 10, 2019
Publication date	Mar 11, 2021
Grant date	—

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

1 . A composition for separating one or more small molecules from a sample comprising:
a size exclusion support; and at least one moiety that can associate with the one or more small molecules.

2 . The composition of claim 1 , wherein the at least one moiety is associated with the size exclusion support or immobilized onto the size exclusion support.

3 . (canceled)

4 . The composition of claim 1 , further comprising least a second size exclusion support and at least a second moiety.

5 . The composition of claim 1 , comprising at least two moieties.

6 . The composition of claim 1 , wherein the at least one moiety comprises a polysaccharide, a dextran, a polyethylene glycol polymer, an amine-containing polymer, a polyaminoacids, a lipopolysaccharide, an antibiotic, a chelating group, a magnetic particle, a paramagnetic particle, a functional group, an ion-exchanger and combinations thereof.

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

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

9 . The composition of claim 6 , wherein the at least one moiety is a dextran.

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

11 . The composition of claim 1 , wherein the at least one 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.

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

13 . The composition of claim 1 , wherein the one or more small molecules comprise a dye, a derivative of a dye, biotin, a biotin derivative, a crosslinker, a reducing agent, a label, a nanoparticle, a radioactive ligand, a mass tag, an unreacted molecule and combinations, intermediates and derivatives thereof.

14 . The composition of claim 1 , wherein the size exclusion supports comprises a dextran polymer, agarose, polyacrylamide, a cellulose material and/or derivatives thereof, a hydroxyethylcellulose or a derivative of a hydroxyethyl cellulose.

15 . 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 quantity of the one or more small molecules from the sample.

16 . A device for separating one or more small molecules from a sample comprising:
a) a container comprising: at least one size exclusion support and at least one moiety that can associate with the one or more small molecules; and b) a receptacle located below the container, wherein the at least one moiety is immobilized onto, attached to, or associated with the size exclusion resin.

17 . The device of claim 16 , operably configured to be subject to a gravity flow, a centrifugal force, a positive pressure, a negative pressure, vacuum and combinations thereof.

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

19 .- 35 . (canceled)

36 . A system for separating one or more small molecules from a sample comprising:
a) a container comprising: a size exclusion support and at least one moiety that can associate with the one or more small molecules; 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 and combinations thereof.

37 . A kit for separating a biomolecule from one or more small molecules comprising:
a device of claim 16 , wherein the device is operably configured to be subject to a gravity flow, a centrifugal force, a positive pressure, a negative pressure, vacuum and combinations thereof.

38 .- 40 . (canceled)

41 . A method for separating a biomolecule from one or more small molecules comprising:
a) applying a sample comprising the biomolecule to a container comprising a size exclusion resin and at least one moiety that can bind to the one or more small molecules; and b) subjecting the container to a gravity flow, a centrifugal force, a positive pressure, a negative pressure, a vacuum or a combination thereof, wherein the biomolecule in the sample is excluded through the size exclusion resin and is collected as a flow through, and wherein the one or more small molecules associates with the at least one moiety and is thereby separated from the biomolecule.

42 .- 43 . (canceled)

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

• C07K1/14

  Extraction; Separation; Purification · CPC title

• B01J20/3042

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