Cell- or virus simulating means comprising encapsulated marker molecules

• US 10,221,445 B2
• Filed: 08/13/2012
• Issued: 03/05/2019
• Est. Priority Date: 08/11/2011
• Status: Active Grant

• Current Assignee: Qiagen GmbH (Qiagen NV)
• Original Assignee: Qiagen GmbH (Qiagen NV), Qiagen Hamburg GmbH (Qiagen NV)
• Inventors: Beller, Katharina, Doedt, Thomas, Heckel, Dirk, Soller, Rainer
• Primary Examiner(s): Chestnut, Barry A
• Application Number:
  US14/238,301
• Publication Number:
  US 20140248606A1
• Time in Patent Office: 2,395 Days
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6811 Selection methods for production or design of target specific oligonucleotides or binding molecules
• CPC Class Codes: C12Q 1/6848 characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction
• CPC Class Codes: C12Q 2545/113 with an external standard/control, i.e. control reaction is separated from the test/target reaction
• CPC Class Codes: C12Q 2547/107 Use of permeable barriers, e.g. waxes
• CPC Class Codes: C12Q 2563/149 Particles, e.g. beads
• CPC Class Codes: C12Q 2563/179 the label being a nucleic acid

Abstract

The present invention refers to a method, a composition and a kit for isolating biomolecules from any biological sample material containing cells, virus(es), microorganism(s) or a combination thereof comprising a cell- or virus-simulating means, wherein said cell- or virus-simulating means comprises at least one type of marker molecule(s), incorporated in at least one type of a layer, capsule, bead, sphere or particle, which is not a biological cell or provided on a substrate covered by a coating.

Claims

1. A cell- or virus-simulating artificial, solid object or article, comprising:

a core and a shell,

wherein the core comprises at least one type of marker molecules selected from the group consisting of nucleic acids, oligopeptides, polypeptides, proteins, biological cells, biological organisms, viruses, phages, or phage core particles, and wherein the shell covers and encloses all of the at least one type of molecules, and

wherein the shell does not comprise a polypeptide.

2. The cell- or virus-simulating artificial, solid object or article according to claim 1, wherein the at least one type of marker molecules is selected from nucleic acids, oligopeptides, polypeptides, and proteins.

3. The cell- or virus-simulating artificial, solid object or article according to claim 1, wherein the at least one type of marker molecules is DNA having a pre-known sequence of at least 10 bp.

4. The cell- or virus-simulating artificial, solid object or article according to claim 1, wherein the cell- or virus-simulating artificial, solid object or article is on an inner surface of a device,

the at least one type of marker molecules is applied on said inner surface, and

a coating or layer covers all of the marker molecule(s) on the inner surface.

5. The cell- or virus-simulating artificial, solid object or article according to claim 1, wherein the cell- or virus-simulating artificial, solid object or article is represented by a bead, capsule, sphere or particle providing at least one of the following features:

(i) size of 10 nm to 1 mm, 100 nm to 100 μ

m, 0.5 to 50 μ

m, or 1 to 10 μ

m;

(ii) the core further comprises a liquid or an inorganic or organic template, and is liquid, solid or porous; and

(iii) the coating or shell covering or the marker molecules comprises an assembling material selected from polyelectrolytes, organic polymers, polypeptides, and phospholipids.

6. A method for isolating and/or analysing biomolecules from a virus-, cell- or microorganism(s)-containing biological sample, comprising:

lysing the biological sample in the presence of or in parallel with the cell- or virus-simulating artificial, solid object or article according to claim 1.

7. A method for the control of a deliberate release of an encapsulated or embedded marker molecule comprised in the cell- or virus-simulating artificial, solid object or article of claim 1, comprising:

lysing the cell- or virus-simulating artificial, solid object or article, and

detecting the at least one type of marker molecules.

8. The method according to claim 7, wherein the method is used for finding, defining, establishing and/or controlling the conditions suitable for effective biomolecule release from a biological sample.

9. The method according to claim 8, wherein the method is used for finding, defining, establishing and/or controlling the conditions suitable for effective biomolecule release from a biological sample in a biomolecule release or isolation process.

10. A method for the control and/or estimation of the success and/or extent of breakage or lysis of a biological cell material, virus or microorganism(s), comprising:

lysing the biological cell material, virus or microorganism(s) in the presence of or in parallel with the cell- or virus-simulating artificial, solid object or article according to claim 1, and

detecting the marker molecule.

11. A method for deliberate release of a marker molecule or a biomolecule into a liquid sample, comprising:

chemically or mechanically lysing the cell- or virus-simulating artificial, solid object or article of claim 1 in a liquid sample, thereby releasing the marker molecule or the biomolecule into the liquid sample.

12. The method according to claim 6, wherein the cell- or virus-simulating artificial, solid object or article

is added to the biological sample,

is present in a device used during the isolation of the biomolecules from the biological sample, or

is used separately from the biological sample in a parallel process.

13. The method according to claim 12, wherein the cell- or virus-simulating artificial, solid object or article

is added to the biological sample,

is present in a device used during the isolation of the biomolecules from the biological sample, or

is used separately from the biological sample in a parallel process that is carried out in the same way.

14. The method according to claim 6, wherein the marker molecules of the cell- or virus-simulating artificial, solid object or article are released during the isolation of the biomolecules from the biological sample, and are isolated together or in parallel with the biomolecules of the biological sample.

15. A composition for lysis of a biological cell material, virus or microorganism comprising the cell- or virus-simulating artificial, solid object or article according to claim 1.

16. The composition according to claim 15, further comprising at least one component selected from water, buffering substance(s), a chaotropic agent, and a detergent.

17. An isolation device for use in a biomolecule isolation method, comprising:

on at least one inner surface coming in contact with a biological cell material, virus or microorganism(s) during the isolation method, a cell- or virus-simulating artificial, solid object or article of claim 1.

18. The isolation device according to claim 17, wherein the at least one type of marker molecules is labelled by a dye, a fluorescent dye, a dye developing compound, an antigen, an antibody, green fluorescent protein (GFP), or radioactivity.

19. The isolation device according to claim 17, wherein the at least one of the inner surface(s) is at least partially coated with a coating comprising a first layer comprising the marker molecule(s) and at least one further layer covering the marker molecule(s).

20. The isolation device according to claim 19, wherein said device is selected from a cup, a tube, a column, a microtiter plate, a multiwell plate, and cups/tubes prefilled with buffers and/ or glass beads.

21. A kit for isolating biomolecules from a biological sample containing cells, virus(es) or a microorganism, comprising a cell- or virus-simulating artificial, solid object or article of claim 1.

22. The kit according to claim 21, further comprising at least one of

(a) a lysis buffer,

(b) a mechanical lysis device, and

(c) a tool for detection of the marker molecule.

23. The cell- or virus-simulating artificial, solid object or article of claim 1, wherein the at least one type of marker molecules is selected from biological cells, biological organisms, viruses, and phages or phage core particles.

24. The cell- or virus-simulating artificial, solid object or article of claim 1, wherein the at least one type of marker molecules is labeled by a dye, a fluorescent dye, a dye-developing compound, green fluorescent protein (GFP), radioactivity, an antigen, or an antibody.

25. The cell- or virus-simulating artificial, solid object or article of claim 24, wherein the at least one type of marker molecules is linked to green fluorescent protein (GFP).

26. The cell- or virus-simulating artificial, solid object or article of claim 1, wherein the at least one type of marker molecules is selected from nucleic acids or polypeptides having a pre-known sequence.

27. The cell- or virus-simulating artificial, solid object or article of claim 1, wherein the shell comprises a poly(beta-amino ester).