Devices and methods for isolating target cells

• US 7,754,155 B2
• Filed: 09/18/2002
• Issued: 07/13/2010
• Est. Priority Date: 03/15/2002
• Status: Expired due to Fees

• Current Assignee: Nexell Therapeutics Inc. (Baxter International Inc.)
• Original Assignee: Nexell Therapeutics Inc. (Baxter International Inc.)
• Inventors: Bernstein, Steve, Ross, Amelia A.
• Primary Examiner(s): Jung; Unsu
• Application Number:
  US10/246,898
• Publication Number:
  US 20030175850A1
• Time in Patent Office: 2,855 Days
• Field of Search: 435/4, 435/6, 435/173.4, 435/173.7, 435/287.6, 435/164, 435/172, 435/514, 435/517, 435/518, 435/524, 435 71- 725, 435/173.5, 4352871-2873, 422/52, 422/55, 422/58, 422/61, 422/73, 422/98, 422/82.01, 422/102, 422/82.05, 356/39, 356/73, 356/315, 600/573, 436/164, 436/172, 436/514, 436/517, 436/518, 436/524
• US Class Current: 422/536
• CPC Class Codes: B01L 2300/069 Absorbents; Gels to retain a fluid
• CPC Class Codes: B01L 2300/0825 Test strips
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/54393 Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
• CPC Class Codes: G01N 33/56966 Animal cells

Abstract

A cell isolating device and method is provided to concentrate or isolate cells with specific characteristics from a mixture of different cell types. One embodiment may comprise two subtypes of antibodies that are directly conjugated to biotin (Ab_b) and conjugated to a fluorescent molecule (Ab_f). The conjugated antibodies (Ab_b+Ab_f) bind to the target cells in a mixed cell suspension. The cell suspension is then passed over an immobilized avidin or streptavidin substrate on a glass microscope slide. The biotinylated target cells adhere to the avidin/streptavidin substrate, while the unbound cells are washed off and collected in a wicking member. Captured cells on the avidin/streptavidin substrate may then be visualized directly using a fluorescent microscope or detected and enumerated via an on-board fluorescent detection device.

Claims

1. A device for isolating and identifying a target cell from a cellular mixture, said device comprising:

a solid substantially planar surface having a bioactive coating applied on a top open planar region of said solid substantially planar surface; and

,

at least one bifunctional compound capable of binding to said target cell and to said bioactive coating; and

a delivery conduit mounted on said solid substantially planar surface and communicating said bifunctional compound with said bioactive coating on said open planar region of said solid substantially planar surface;

said delivery conduit mounted on said solid substantially planar surface so as to form an unsealed space between said delivery conduit and said solid substantially planar surface, said unsealed space having a predetermined height and said space allowing flow of said bifunctional compound to travel beyond said delivery conduit across said substantially planar surface over said bioactive coating on said substantially planar surface.

2. The device of claim 1 further comprising a binder molecule conjugated with a visual indicator probe.

3. The device of claim 1 wherein said bioactive coating is selected from the group consisting of avidin, streptavidin, avidin derivatives, streptavidin derivatives, and blends thereof.

4. The device of claim 1 wherein said bifunctional molecule comprises a binding molecule conjugated with a biotin molecule or a biotin derivative.

5. The device of claim 4 wherein said binding molecule is selected from the group consisting of antibodies, antigens, glycoconjugates, lectins, hormones, cell receptors, vitamins, amino acids, sugars, lipids, fatty acids, liposomes, DNA probes, and RNA probes.

6. The device of claim 2 wherein said probe is selected from the group consisting of flurochromes, radiolabels, fluorescent agents, and chromophores.

7. The device of claim 1 wherein said delivery conduit includes a capillary annulus in spaced relationship with said solid substantially planar surface.

8. The device of claim 7 further comprising a fluid absorbing media surrounding said capillary annulus.

9. The device of claim 8 further comprising a specimen chamber in communication with said capillary annulus.

10. The device of claim 1 wherein said delivery conduit includes a housing in spaced relationship with said solid substantially planar surface, said housing having an inlet located at a first end of said housing.

11. The device of claim 10 further comprising a fluid, absorbing media opposite said inlet and adjacent to said bioactive coating.

12. The device of claim 1, wherein said predetermined height of said unsealed space is in the range of approximately 2 to 20 microns.

13. The device of claim 1, further comprising at least one fastener securing said delivery conduit to said substantially planar surface.

14. The device of claim 1, wherein said substantially planar surface is a slide.

15. The device of claim 1, wherein said delivery conduit has a constant diameter.

16. The device of claim 1, wherein said delivery conduit has a tapered shape.

17. The device of claim 1, wherein said delivery conduit has a top wall and a first side wall and a second side wall.