# US 11,305,283 B2 · Micro-fluidic devices for assaying biological activity

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- Number: US 11,305,283 B2
- Document ID: US11305283B2
- Status: Granted
- Kind: B2
- Application number: 16/509,166
- Filed: 2019-07-11
- Published: 2020-03-12
- Granted: 2022-04-19
- Earliest claimed: 2013-10-22
- Assignees: Berkeley Lights, Inc.
- Applicants: Berkeley Lights, Inc.
- Inventors: Kevin T. Chapman; Daniele Malleo; J. Tanner Nevill; Steven W. Short; Mark P. White; M. Jimena Loureiro
- Web page: https://patentgrep.com/patent/US11305283B2

## CPC

- B01L 3/502761: {specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads or physically stretching molecules}
- B03C 5/005: {Dielectrophoresis, i.e. dielectric particles migrating towards the region of highest field strength}
- B03C 5/026: {using open-gradient differential dielectric separation, i.e. using electrodes of special shapes for non-uniform field creation, e.g. Fluid Integrated Circuit [FIC]}
- G01N 15/1484: {microstructural devices}
- G01N 33/5023: {on expression patterns}
- G01N 33/5047: {Cells of the immune system}
- G01N 33/505: {involving T-cells}
- G01N 33/5052: {involving B-cells}
- G01N 33/54313: {the carrier being characterised by its particulate form}
- G01N 33/6854: {Immunoglobulins}
- B01L 2200/0647: Handling flowable solids, e.g. microscopic beads, cells, particles
- B01L 2200/0668: Trapping microscopic beads
- B01L 2300/0636: Integrated biosensor, microarrays
- B01L 2300/0816: Cards, e.g. flat sample carriers usually with flow in two horizontal directions
- B01L 2300/0864: comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
- B01L 2300/087: Multiple sequential chambers
- B01L 2400/0454: radiation pressure, optical tweezers
- B03C 2201/26: for use in medical or biological applications
- B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B03C: MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS (separating isotopes B01D59/00; combinations of magnetic or electrostatic separation with separation of solids by other means B03B, B07B)
- 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)

## Abstract

Biological activity in holding pens in a micro-fluidic device can be assayed by placing in the holding pens capture objects that bind a particular material of interest produced by the biological activity. The biological material of interest that binds to each capture object can then be assessed, either in the micro-fluidic device or after exporting the capture object from the micro-fluidic device. The assessment can be utilized to characterize the biological activity in each holding pen. The biological activity can be production of the biological material of interest. Thus, the biological activity can correspond to or arise from one or more biological cells. Biological cells within a holding pen can be clonal cell colonies. The biological activity of each clonal cell colony can be assayed while maintaining the clonal status of each colony.

## Claims

### Claim 1

A micro-fluidic device comprising a housing wherein said housing comprises:
a base;
a micro-fluidic structure disposed on said base, wherein said micro-fluidic structure and said base define a flow region comprising a channel,
a holding pen disposed on said base and disposed within said flow region, and
an assay region comprising an assay chamber having an opening to said channel, wherein said assay chamber is located beside said holding pen,
wherein said holding pen comprises an isolation region with a single opening, and a connection region, said connection region having a proximal opening to said channel and a distal opening to said isolation region; and
wherein said assay region is located adjacent to said holding pen.

### Claim 2 (depends on 1)

The micro-fluidic device of claim 1, wherein said assay chamber substantially lacks an isolation region.

### Claim 3 (depends on 1)

The micro-fluidic device claim 1, wherein said device further comprises a means for generating a magnetic force within said microfluidic device.

### Claim 4 (depends on 1)

The microfluidic device of claim 1, wherein said housing further comprises:
a first electrode;
a second electrode; and
an electrode activation substrate,
wherein said first electrode is part of a first wall of said housing and said second electrode and said electrode activation substrate are part of a second wall of said housing,
wherein said electrode activation substrate has a surface comprising a plurality of DEP electrode regions, and
wherein said surface of said electrode activation substrate is an inner surface of said flow region.

### Claim 5 (depends on 4)

The microfluidic device of claim 4, wherein said electrode activation substrate is a photoconductive material.

### Claim 6 (depends on 4)

The microfluidic device of claim 4, wherein said electrode activation substrate comprises a semiconductor material.

### Claim 7 (depends on 6)

The microfluidic device of claim 6, wherein said semiconductor material comprises a plurality of doped layers, electrically insulating layers, and electrically conductive layers that form semiconductor integrated circuits.

### Claim 8 (depends on 1)

A process of assaying biological activity in the micro-fluidic device of claim 1, said process comprising:
culturing one or more biological cells in a holding pen of a micro-fluidic device, wherein said one or more cells produce n different biological materials of interest;
introducing n different types of capture micro-objects into said holding pen, each said type of capture micro-object comprising a binding substance that specifically binds to one of said n different biological materials of interest;
allowing said n different biological materials of interest produced by said one or more biological cells to bind to said n different types of capture micro-objects; and assessing binding between said n different biological materials of interest and said n different types of capture micro-objects.

### Claim 9 (depends on 8)

The process of claim 8, wherein a result of said assessing is positive if at least one of said n different biological materials of interest specifically binds to one of said n different types of capture micro-objects.

### Claim 10 (depends on 8)

The process of claim 8, wherein a result of said assessing is positive if at least two of said n different biological materials of interest each specifically binds to one of said n different types of capture micro-objects.

### Claim 11 (depends on 8)

The process of claim 8, wherein a result of said assessing is positive if all n different biological materials of interest each specifically binds to one of said n different types of capture micro-objects.

### Claim 12 (depends on 8)

The process of claim 8, wherein said n different types of capture micro-objects are introduced into said pen simultaneously.

### Claim 13 (depends on 8)

The process of claim 8, wherein said n different types of capture micro-objects are introduced into said pen sequentially.

### Claim 14

A micro-fluidic device comprising a housing wherein said housing comprises:
a base;
a micro-fluidic structure disposed on said base, wherein said micro-fluidic structure and said base define a flow region comprising a channel, a holding pen disposed on said base and disposed within said flow region, and an assay region comprising an assay chamber having an opening to said channel,
wherein said holding pen comprises an isolation region with a single opening, and a connection region, said connection region having a proximal opening to said channel and a distal opening to said isolation region;
wherein said assay region is located adjacent to said holding pen, and wherein said opening of said assay chamber is located directly across said channel from said proximal opening of said connection region of said holding pen.

### Claim 15 (depends on 14)

The micro-fluidic device of claim 14, wherein said assay chamber substantially lacks an isolation region.

### Claim 16 (depends on 14)

The micro-fluidic device claim 14, wherein said device further comprises a means for generating a magnetic force within said microfluidic device.

### Claim 17 (depends on 14)

The microfluidic device of claim 14, wherein said housing further comprises:
a first electrode;
a second electrode; and
an electrode activation substrate,
wherein said first electrode is part of a first wall of said housing and said second electrode and said electrode activation substrate are part of a second wall of said housing,
wherein said electrode activation substrate has a surface comprising a plurality of DEP electrode regions, and
wherein said surface of said electrode activation substrate is an inner surface of said flow region.

### Claim 18 (depends on 17)

The microfluidic device of claim 17, wherein said electrode activation substrate is a photoconductive material.

### Claim 19 (depends on 17)

The microfluidic device of claim 17, wherein said electrode activation substrate comprises a semiconductor material.

### Claim 20 (depends on 19)

The microfluidic device of claim 19, wherein said semiconductor material comprises a plurality of doped layers, electrically insulating layers, and electrically conductive layers that form semiconductor integrated circuits.