# US 7,488,603 B2 · Method and device for extracting an analyte

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- Number: US 7,488,603 B2
- Document ID: US7488603B2
- Status: Granted
- Kind: B2
- Application number: 10/920,960
- Filed: 2004-08-17
- Published: 2005-01-27
- Granted: 2009-02-10
- Earliest claimed: 2004-05-12
- Assignees: PhyNexus, Inc.
- Inventors: Douglas T. Gjerde; Christopher P. Hanna
- Web page: https://patentgrep.com/patent/US7488603B2

## CPC

- B01L 3/0275: {Interchangeable or disposable dispensing tips}
- B01D 15/3804: {Affinity chromatography}
- B01J 20/285: based on polymers
- B01J 20/291: Gel sorbents
- B82Y 30/00: Nanotechnology for materials or surface science, e.g. nanocomposites
- G01N 35/10: Devices for transferring samples {or any liquids} to, in, or from, the analysis apparatus, e.g. suction devices, injection devices {(G01N35/0099 takes precedence)}
- B01D 15/325: {Reversed phase}
- B01D 15/327: {with hydrophobic interaction}
- B01J 2220/64: In a syringe, pipette, e.g. tip or in a tube, e.g. test-tube or u-shape tube
- B01L 2200/0631: Purification arrangements, e.g. solid phase extraction [SPE]
- G01N 30/6004: {end pieces}
- G01N 2030/009: {Extraction}
- G01N 2035/103: {using disposable tips}
- G01N 2035/1039: {Micropipettes, e.g. microcapillary tubes}
- G01N 2035/1053: {for separating part of the liquid, e.g. filters, extraction phase}
- Y10T 436/153333: Halogen containing
- Y10T 436/25375: Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]
- Y10T 436/255: including use of a solid sorbent, semipermeable membrane, or liquid extraction
- Y10T 436/2575: Volumetric liquid transfer
- B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01D: SEPARATION (separating solids from solids by wet methods B03B, B03D; by pneumatic jigs or tables B03B; by other dry methods B07; magnetic or electrostatic separation of solid materials from solid materials or fluids, separation by high-voltage electric fields B03C; centrifuges, vortex apparatus B04; presses per se for squeezing-out liquid from liquid-containing material B30B9/02; treatment of water C02F, e.g. softening by ion-exchange C02F1/42; {arrangements of air intake cleaners in gas turbine plants F02C7/05} ; arrangements or mounting of filters in air-conditioning, air-humidification or ventilation F24F13/28)
- B01J: CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B82Y: SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- 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)
- Y10T: TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION

## Abstract

The invention provides extraction columns for the purification of an analyte (e.g., a biological macromolecule, such as a peptide, protein or nucleic acid) from a sample solution, as well as methods for making and using such columns. The columns typically include a bed of extraction media positioned in the column between two frits. In some embodiments, the extraction columns employ modified pipette tips as column bodies. In some embodiments, the invention provides methods characterized by the elution of analyte in a small volume of liquid.

## Claims

### Claim 1

A method of extracting an analyte from a solution comprising the steps of:
a. passing a solution containing an analyte back and forth through a pipette tip column, wherein the pipette tip column is comprised of:
i. a column body having an open upper end, an open lower end, and an open channel between the upper and lower ends of the column body,
ii. a bottom frit bonded to and extending across the open channel,
iii. a top frit bonded to and extending across the open channel between the bottom frit and the open upper end of the column body, wherein the top frit, bottom frit, and column body define the extraction media chamber, wherein the top frit and the bottom frit are less than 350 microns thick, wherein the bottom frit is a membrane screen and the top frit is optionally a membrane screen, and wherein the membrane screen is selected from a group of materials consisting of nylon, polyester, polyamide, polycarbonate, cellulose, polyethylene, nitrocellulose, cellulose acetate, polypropylene, polysulfone, and glass, and
iv. a bed of extraction media having a bed volume, whereby analyte adsorbs to said extraction media; and
b. eluting the analyte by aspirating and discharging a desorption solvent through the pipette tip column, and wherein said analyte is an active protein.

### Claim 2 (depends on 1)

The method of claim 1, wherein the volume of desorption solvent is in the range of about 10% to about 300% of the bed volume.

### Claim 3 (depends on 1)

The method of claim 1, wherein step (a) is performed more than one time.

### Claim 4 (depends on 1)

The method of claim 1, wherein the bed volume is in the range of between 0.1 and 100 uL.

### Claim 5 (depends on 1)

The method of claim 1, wherein between step (a) and step (b), a quantity of wash solution is aspirated and discharged through the extraction media.

### Claim 6 (depends on 1)

The method of claim 1, wherein step (b) is performed more than one time.

### Claim 7 (depends on 1)

The method of claim 1, wherein the extraction media comprises an affinity binding agent.

### Claim 8 (depends on 1)

The method of claim 1, wherein the bed of extraction media comprises a packed bed of gel resin beads.

### Claim 9 (depends on 8)

the method of claim 8, wherein the gel resin beads are agarose beads or sepharose beads.

### Claim 10 (depends on 1)

The method of claim 1, wherein the bottom frit has a pore volume of 1 microliter or less.

### Claim 11 (depends on 1)

The method of claim 1, wherein the bottom frit is less than 200 microns thick.

### Claim 12 (depends on 1)

The method of claim 1, wherein the upper end of the column body is attached to a pump for aspirating fluid through the lower end of the column body.

### Claim 13 (depends on 12)

The method of claim 12, wherein the pump is a pipettor, a syringe, or a peristaltic pump.