# US 9,428,725 B2 · Compositions and methods for capture and elution of biological materials via particulates

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- Number: US 9,428,725 B2
- Document ID: US9428725B2
- Status: Granted
- Kind: B2
- Application number: 14/155,123
- Filed: 2014-01-14
- Published: 2014-05-08
- Granted: 2016-08-30
- Earliest claimed: 2010-12-23
- Assignees: Claremont Biosolutions LLC
- Applicants: Claremont BioSolutions, LLC
- Inventors: Mark Brown; Robert Doebler; Bruce Irvine; Tanya Ferguson; Gary Blackburn
- Web page: https://patentgrep.com/patent/US9428725B2

## CPC

- C12N 1/06: Lysis of microorganisms
- C12M 47/06: {Hydrolysis; Cell lysis; Extraction of intracellular or cell wall material (lysis of microorganisms C12N1/06; extracting or separating nucleic acids from biological samples C12N15/1003)}
- G01N 33/54313: {the carrier being characterised by its particulate form}
- B01L 3/563: {Joints or fittings; Separable fluid transfer means to transfer fluids between at least two containers, e.g. connectors}
- B01L 3/567: {Valves, taps or stop-cocks}
- G01N 2001/2866: {Grinding or homogeneising}
- G01N 2001/4088: {filtration}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12M: APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; {APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS}
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE

## Abstract

A flow-through device for capturing a biological material from a fluid specimen and lysing the fluid specimen. The flow through device may include a first chamber containing a portion of the fluid specimen and a solid phase material retained in the first chamber. The solid phase material may have an affinity for the biological material in the fluid specimen. The first chamber may include an inlet opening which allows flow of the fluid specimen into the first chamber. The first chamber may also include an outlet opening which allows flow of the fluid specimen out of the first chamber. The flow through device may include a second chamber fluidly connected to the first chamber. The second chamber may include a portion of the fluid specimen and an impeller. The impeller may in use impart rotational motion to the fluid specimen to mechanically lyse the fluid specimen.

## Claims

### Claim 1

A flow-through device for capturing a biological material from a fluid specimen and lysing the fluid specimen, the device comprising:
a first chamber containing:
a portion of the fluid specimen;
a solid phase material retained in the first chamber, the solid phase material having an affinity for the biological material in the fluid specimen;
wherein the first chamber comprises:
an inlet opening allowing flow of the fluid specimen into the first chamber;
a first fitting attached to the inlet opening of the first chamber, the first fitting having a first thin filter insert that retains the solid phase material while allowing rapid flow of the fluid specimen at low pressure;
an outlet opening allowing flow of the fluid specimen out of the first chamber; and
a second fitting attached to the outlet opening of the first chamber, the second fitting having a second thin filter insert that retains the solid phase material while allowing rapid flow of the fluid specimen at low pressure; and
a second chamber fluidly coupled to the first chamber, the second chamber containing:
a portion of the fluid specimen;
wherein the second chamber comprises:
an impeller which in use imparts rotational motion to the fluid specimen to mechanically lyse the fluid specimen.

### Claim 2 (depends on 1)

The device of claim 1 wherein the second chamber is connected to the inlet opening of the first chamber.

### Claim 3 (depends on 1)

The device of claim 1 wherein the second chamber is connected to the outlet opening of the first chamber.

### Claim 4 (depends on 1)

The device of claim 1 wherein the solid phase material has an affinity for at least one selected from the group consisting of a nucleic acid, a protein, a polypeptide, a His-tagged protein, a His-tagged polypeptide, a GST-tagged protein, a GST-tagged polypeptide, streptavidin, biotin, calmodulin, an antigen, an antibody, an immunoglobulin, albumin, DNA, RNA, double stranded DNA, an RNA/DNA heteroduplex, maltose binding protein, FLAG-tagged protein, FLAG-tagged polypeptide, MYC-tagged protein, MYC-tagged polypeptide MYC, a glycoprotein, a SNAP-tag, an enzyme, a lipid-containing biological material, a glycosylated protein, a phosphorylated protein, and a microorganism having a high cell wall lipid content.

### Claim 5 (depends on 1)

The device of claim 1 wherein the solid phase material has an affinity for more than one biological material.

### Claim 6 (depends on 1)

The device of claim 1 wherein the solid phase material is a particulate or a bead.

### Claim 7 (depends on 6)

The device of claim 6 wherein the particulate or bead has a diameter or lateral dimension of at least 10 μm.

### Claim 8 (depends on 1)

The device of claim 1 wherein the solid phase material comprises at least one of a plurality of ceramic beads, a plurality of glass beads, a plurality of zirconia beads, a plurality of gold beads, a plurality of gold-coated beads, a plurality of silica beads, a plurality of sand, a plurality of beads with a metal core coated by a material that facilitates binding of the biological material, a plurality of mineral oxide beads coated with silane compounds to impart hydrophobicity and/or positive charge, and a plurality of paramagnetic beads.

### Claim 9 (depends on 1)

The device of claim 1 wherein the first and second thin filter inserts have a pore size between about 10 μm and about 200 μm.

### Claim 10 (depends on 1)

The device of claim 1 wherein the first and second thin filter inserts are wire mesh inserts.

### Claim 11 (depends on 1)

The device of claim 1 wherein the first and second thin filter inserts are plastic mesh inserts.

### Claim 12

A method of capturing a biological material from a fluid specimen and lysing the fluid specimen, the method comprising:
introducing the fluid specimen containing the biological material into a flow-through device comprising a first chamber, the first chamber containing a solid phase material retained in the first chamber, the solid phase material having an affinity for the biological material; the first chamber having an inlet opening and an outlet opening, the inlet opening and the outlet opening allowing flow of the fluid specimen into and out of the first chamber, the first chamber further having a first fitting attached to the inlet opening of the first chamber, the first fitting having a first thin filter insert that retains the solid phase material while allowing rapid flow of the fluid specimen at low pressure, the first chamber further having a second fitting attached to the outlet opening of the first chamber, the second fitting having a second thin filter insert that retains the solid phase material while allowing rapid flow of the fluid specimen at low pressure; flowing the fluid specimen containing the biological material through the solid phase material, the fluid specimen exiting the first chamber via the outlet opening; and lysing the fluid specimen in a second chamber fluidly connected to the first chamber, the second chamber comprising: an impeller which in use imparts rotational motion to the fluid specimen to mechanically lyse the fluid specimen.

### Claim 13 (depends on 12)

The method of claim 12 wherein lysing the fluid specimen comprises agitating the specimen in the second chamber with a medium that includes a particulate material to mechanically lyse the specimen.

### Claim 14 (depends on 12)

The method of claim 12 wherein introducing the fluid specimen into the first chamber of a device includes introducing the specimen into the first chamber containing a solid phase material having an affinity for at least one selected from the group consisting of a nucleic acid, a protein, a polypeptide, a His-tagged protein, a His-tagged polypeptide, a GST-tagged protein, a GST-tagged polypeptide, streptavidin, biotin, calmodulin, an antigen, an antibody, an immunoglobulin, albumin, DNA, RNA, double stranded DNA, an RNA/DNA heteroduplex, maltose binding protein, FLAG-tagged protein, FLAG-tagged polypeptide, MYC-tagged protein, MYC-tagged polypeptide MYC, a glycoprotein, a SNAP-tag, an enzyme, a lipid-containing biological material, a glycosylated protein, a phosphorylated protein, and a microorganism having a high cell wall lipid content.

### Claim 15 (depends on 12)

The method of claim 12 wherein introducing the fluid specimen into the first chamber of a device includes introducing the specimen into the first chamber containing a solid phase material having an affinity for more than one biological material.

### Claim 16 (depends on 12)

The method of claim 12 wherein introducing the fluid specimen into the first chamber of a device includes introducing the specimen into the first chamber containing a solid phase material comprising a particulate or a bead.

### Claim 17 (depends on 16)

The method of claim 16 wherein introducing the fluid specimen into the first chamber of the device includes introducing the specimen into the first chamber containing a solid phase material comprising a particulate or bead having a diameter or lateral dimension of at least 10 μm.

### Claim 18 (depends on 12)

The method of claim 12 wherein the first and second thin filter inserts have a pore size between about 10 μm and about 200 μm.

### Claim 19 (depends on 12)

The method of claim 12 wherein introducing the fluid specimen into the first chamber includes introducing the specimen into the first chamber, and the first and second thin filter inserts comprise wire mesh inserts.

### Claim 20 (depends on 12)

The method of claim 12 wherein introducing the fluid specimen into the first chamber includes introducing the specimen into the first chamber, and the first and second thin filter inserts comprise stainless steel mesh inserts.

### Claim 21 (depends on 12)

The method of claim 12 wherein introducing the fluid specimen into the first chamber includes introducing the specimen into the first chamber, and the first and second thin filter inserts comprise plastic mesh inserts.

### Claim 22 (depends on 12)

The method of claim 12, further comprising:
eluting the biological material captured on the solid phase material by flowing an elution medium through the solid phase material; and collecting an effluent containing the biological material, the effluent exiting the device.