# US 10,369,568 B2 · Cell separation using microchannel having patterned posts

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- Number: US 10,369,568 B2
- Document ID: US10369568B2
- Status: Granted
- Kind: B2
- Application number: 15/193,712
- Filed: 2016-06-27
- Published: 2017-05-25
- Granted: 2019-08-06
- Earliest claimed: 2005-01-18
- Assignees: Biocept, Inc.
- Applicants: Biocept, Inc.
- Inventors: Zhongliang Tang; Ram Bhatt; Pavel Tsinberg
- Web page: https://patentgrep.com/patent/US10369568B2

## CPC

- B01L 3/502753: {characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation}
- F04D 19/04: specially adapted to the production of a high vacuum, e.g. molecular pumps
- F04D 19/042: {Turbomolecular vacuum pumps}
- G01N 1/34: Purifying; Cleaning {(processes or apparatus for extracting or separating nucleic acids from biological samples C12N15/1003)}
- G01N 33/491: {by separating the blood components (G01N15/05 takes precedence)}
- G01N 33/54386: {Analytical elements}
- B01L 2200/0647: Handling flowable solids, e.g. microscopic beads, cells, particles
- B01L 2200/12: Specific details about manufacturing devices
- B01L 2200/141: Preventing contamination, tampering
- B01L 2300/0681: Filter
- B01L 2300/0816: Cards, e.g. flat sample carriers usually with flow in two horizontal directions
- B01L 2300/0825: Test strips
- B01L 2300/16: Surface properties and coatings
- B01L 2400/0457: passive flow or gravitation
- B01L 2400/086: using baffles or other fixed flow obstructions
- F16C 2360/45: Turbo-molecular pumps
- B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- F04D: NON-POSITIVE-DISPLACEMENT PUMPS (engine fuel-injection pumps F02M; ion pumps H01J41/12; electrodynamic pumps H02K44/02)
- 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)
- F16C: SHAFTS; FLEXIBLE SHAFTS; ELEMENTS OR CRANKSHAFT MECHANISMS; ROTARY BODIES OTHER THAN GEARING ELEMENTS; BEARINGS

## Abstract

A micro flow device (11, 71) for separating or isolating cells from a bodily fluid or other liquid sample uses a flow path where straight-line flow is interrupted by a pattern of transverse posts (23, 81). The posts are spaced across the width of an expanded collection chamber region (17, 75) in the flow path, extending between the upper and lower surfaces thereof; they have rectilinear surfaces, being curved in cross-sections, e.g. circular or tear-drop shaped, and are randomly arranged so as to disrupt streamlined flow. The device is oriented so that its lower surface is aligned at about 45° to the horizontal. Sequestering agents, such as Abs, which are attached to surfaces of the collection region via a hydrophilic coating, preferably a permeable hydrogel containing isocyanate moieties, are highly effective in capturing cells or other targeted biomolecules while the remainder of the liquid sample exits horizontally.

## Claims

### Claim 1

A method for separating biomolecules, such as cells, from a sample of bodily fluid or other liquid, which method comprises:
causing such a sample containing target biomolecules to flow downstream along a flow path in a microflow device from an inlet to an outlet, which flow path comprises a microchannel arrangement that includes a collection region of expanded cross section, while orienting the device so that said flow path collection region is aligned at an angle of about 30° to about 60° to the horizontal, wherein the sample exits the device through the outlet in a substantially horizontal direction,
separating target biomolecules from the flowing sample by
(a) interrupting straight-line flow of the liquid through said collection region, as a result of blocking such flow with a plurality of separator posts located in said region, which posts are integral with an upper or lower surface of said microchannel and extend therefrom to the opposite surface thereof, said posts extending transverse to said flow path and being located in an irregular pattern that extends laterally across said collection region and prevents straight-line flow and streamlined flow therethrough, and all surfaces of said collection region including said posts having sequestering agents carried thereupon, and
(b) capturing target biomolecules found in the flowing liquid sample on surfaces in the collection region by binding the target molecules to the sequestering agents as a result of flow disruption by said irregular posts and force vectors that result from gravity, which vectors are aligned at an acute angle to said lower surface of said collection region, and
discharging the remainder of the liquid sample through the outlet.

### Claim 2 (depends on 1)

The method according to claim 1 wherein the sample is fed into a well at the inlet and drawn through said channel and out the outlet by vacuum.

### Claim 3 (depends on 1)

The method according to claim 1 wherein said device includes (a) a body having a flat surface wherein the collection region is formed as a cavity and (b) a flat plate which abuts said flat surface and forms a bottom wall.

### Claim 4 (depends on 3)

The method according to claim 3 wherein said flat plate is optically transparent and wherein optical detection means which is positioned thereadjacent monitors parameters of biomolecules captured in said collection region and provides an output signal corresponding to a monitored parameter.

### Claim 5 (depends on 1)

The method according to claim 1 wherein said inlet passageway is aligned at an angle of about 40° to 50° to the flow path through the collection region.

### Claim 6 (depends on 5)

The method according to claim 5 wherein said outlet passageway lies in substantially the same vertical plane as said inlet passageway and is oriented at an angle of about 90° to said inlet passageway in said vertical plane and wherein exit therethrough is in a substantially horizontal direction.

### Claim 7 (depends on 1)

The method according to claim 1 wherein said sample is caused to flow through said collection region at an average liquid flow rate of about 0.2 to about 1 mm/sec.

### Claim 8 (depends on 1)

The method according to claim 1 wherein said surfaces in said collection region have a hydrophilic coating, which coating is a permeable hydrogel formed from an isocyanate-functional prepolymer that is a reaction product of PEG, PPG or a copolymer thereof and a polyisocyanate, and wherein said sequestering agents are indirectly bound to said hydrogel coating by coupling pairs.

### Claim 9 (depends on 8)

The method according to claim 8 wherein said surfaces in said collection region are coated in situ by supplying an aqueous mixture containing said prepolymer thereto while said prepolymer undergoes final polymerization.

### Claim 10 (depends on 1)

The method according to claim 1, wherein said surfaces in said collection region are coated with PEG.