Methods of specifically releasing a sub-group of objects

Patent metadata

Field	Value
Publication number	US-9506918-B2
Application number	US-201514673049-A
Country	US
Kind code	B2
Filing date	Mar 30, 2015
Priority date	Oct 2, 2012
Publication date	Nov 29, 2016
Grant date	Nov 29, 2016

Abstract

The present invention relates to a method of specifically releasing a one or more members of a sub-group of objects from an entity, a method of detecting a subject's disease by detecting one or more members of a sub-group of biological entities indicative of the disease and a method of isolating one or more members of a sub-group of objects from a group of objects.

First claim

The invention claimed is:

1. A method of specifically releasing one or more members of a sub-group of objects from an entity, the method comprising
a) providing a heterogeneous group of objects comprising a homogeneous subgroup of objects, each member of the group of objects being bound to the entity via a linker;
b) contacting the group of objects with an identifier, being capable of specifically binding the subgroup of objects, under conditions allowing the binding of the identifier to the subgroup of objects, wherein the identifier is coupled to a releaser being capable of releasing a member of the subgroup from the entity by interacting with the linker;
c) binding the identifier to the subgroup, thereby bringing the releaser in proximity to the linker; and
d) specifically releasing one or more members of the subgroup from the entity by allowing interaction between releaser and linker;
thereby specifically releasing the one or more members of the subgroup of objects from the entity.

2. The method of claim 1 , wherein the entity is a support, particularly a solid support.

3. The method of claim 1 , wherein the objects are proteins, nucleic acids, antigens, antibodies or functionally active derivatives thereof.

4. The method of claim 1 , wherein the heterogeneous group of objects comprising a homogeneous subgroup of objects is a heterogeneous population of cells comprising a homogeneous sub-population of cells.

5. The method of claim 4 , wherein
a) the sub-population of cells consists of rare cells, particularly wherein in the population the ratio of rare cells to total cells is at most 5%, preferably at most 1%, especially at most 0.1%, such as at most 0.01%; or
b) in the population the ratio of cells of the homogeneous sub-population of cells to total cells is at most 50%, especially in the range from 5% to 50%, particularly in the range from 10% to 30%.

6. The method of claim 4 , wherein the sub-population of cells is
indicative of a disease, particularly of cancer; or
composed of cells of one cell type.

7. The method of claim 1 , wherein the identifier is selected from the group comprising a protein, a nucleic acid, an antibody or functionally active derivative thereof, a receptor or a binding site thereof, an enzyme or a binding site thereof, a tag-binding site, a ligand to a receptor, a substrate to an enzyme or a tag.

8. The method of claim 1 , wherein the linker comprises
(i) a cleavage site for an enzyme;
(ii) a nucleic acid comprising a cleavage site for an enzyme;
(iii) a protein comprising a cleavage site for an enzyme;
(iv) a singlet oxygen-sensitive bond; or
(v) a pole sensitive to an electronically controllable charged group in the releaser.

9. The method of claim 1 , wherein the releaser is an enzyme, an apoenyzme, or a portion of a split enzyme, a photo sensitizer, an electronically controllable charged group targeting an opposite pole in the linker or a metal complex.

10. The method of claim 1 , wherein the releasing is by cleaving the linker, particularly by
(i) enzyme-mediated cleaving of the linker;
(ii) photochemical, particularly singlet oxygen-mediated, cleaving of the linker;
(iii) chemical, particularly reductive, cleaving of the linker; or
(iv) electrochemical cleaving of the linker.

11. The method of claim 2 , wherein the solid support is
(i) a bead, a vessel, a slide, a disk such as a compact disk or an array such as a microarray, and/or
(ii) glass, polystyrene, polyacrylamide, poly carbonate, polyethylene glycol (PEG), PEG-based or cellulose, optionally having a functionalized surface or being a chemically modified support, and/or
(iii) a petri dish, a microwell plate, a chromatography column, a chromatography matrix, a cell culture flask, a tube, a mesh or a membrane.

12. A method of detecting a subject's disease by detecting a sub-group of biological units indicative of the disease, the method comprising
a) providing a heterogeneous group of biological units comprising a homogeneous subgroup of biological unity, the group of biological units being obtained from a subject and each member of the group of biological units being bound to an entity, preferably a solid support, via a linker; and
b) contacting the group of biological units with a identifier, being capable of specifically binding the subgroup of biological units, under conditions allowing the binding of the identifier to the subgroup of biological units, wherein the identifier is coupled to a releaser, being capable of releasing the subgroup from the entity by interacting with the linker;
c) binding the identifier to the subgroup, thereby bringing the releaser in proximity to the linker;
d) specifically releasing one or more members of the subgroup from the entity by allowing interaction between the releaser and the linker; and
e) detecting the presence of one or more members of the subgroup, thereby detecting the disease.

13. A method of isolating one or more members of a sub-group of objects from a group of objects, the method comprising
a) providing a heterogeneous group of objects comprising a homogeneous subgroup of objects, each member of the group of objects being bound to an entity, preferably a solid support, via a linker; and
b) contacting the group of objects with a identifier, being capable of specifically binding the subgroup of objects, under conditions allowing the binding of the identifier to the subgroup of objects, wherein the identifier is coupled to a releaser being capable of releasing the subgroup from the entity by interacting with the linker;
c) binding the identifier to one or more members of the subgroup, thereby bringing the releaser in proximity to the linker;
d) specifically releasing one or more members of the subgroup from the entity by allowing interaction between the releaser and the linker; and
e) isolating the released one or more members of the subgroup.

Assignees

• Roche Diagnostics Operations Inc

Inventors

• Fischer Thomas
• Froehlich Thomas
• Hendl Dieter

Classifications

• G01N33/5005

  involving human or animal cells (immunoassay G01N33/56966; immunoassays of protozoa G01N33/56905; protozoa in screening assays C12Q1/025) · CPC title

• G01N33/54306 Primary

  Solid-phase reaction mechanisms · CPC title

• C12Q1/6834

  Enzymatic or biochemical coupling of nucleic acids to a solid phase · CPC title

• C12Q1/6823

  Release of bound markers · CPC title

• G01N2800/00

  Detection or diagnosis of diseases · CPC title