Binding method and apparatus for sorting objects

• US 20090068671A1
• Filed: 09/11/2008
• Published: 03/12/2009
• Est. Priority Date: 09/11/2007
• Status: Active Grant

• Current Assignee: Arryx, Inc. (Haemonetics Corporation)
• Original Assignee: Arryx, Inc. (Haemonetics Corporation)
• Inventors: Chakrabarty, Tania
• Granted Patent: US 8,067,170 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: G01N 2800/367 Infertility, e.g. sperm disorder, ovulatory dysfunction
• CPC Class Codes: G01N 33/5091 for testing the pathological state of an organism
• CPC Class Codes: G01N 33/56966 Animal cells

Abstract

The present invention relates to a method and apparatus of sorting objects including, providing a sample having wanted objects and unwanted objects; coating a surface of a sample holder with an antibody; placing an eluted sample on the sample holder; binding an antigen in the wanted objects with the antibody on the surface of the sample holder to sort the objects into wanted objects and unwanted objects; separating the wanted objects; and performing PCR-based STR analysis on the wanted objects. In one embodiment, holographic optical trapping is used to further sort the wanted objects. In other embodiments, the wanted objects are sperm and the antibody is a human sperm specific antibody, and the PCR is single cell PCR-based STR analysis. In still other embodiments, the binding is direct or indirect, ligands are used to bind to object-specific organomolecules, and protein A or protein G are used to bind the antibody.

Claims

1. A method of sorting objects comprising:

providing a sample having wanted objects and unwanted objects;

coating a surface of a sample holder with an antibody;

eluting said sample and placing said eluted sample on said sample holder;

binding an antigen in wanted objects in said sample with said antibody on said surface of said sample holder to sort said objects into wanted objects and unwanted objects;

separating said wanted objects;

removing said unwanted objects; and

performing PCR-based STR analysis on said wanted objects.

2. The method of claim 1, performing holographic optical trapping to sort said wanted objects from said unwanted objects.

3. The method of claim 1, wherein said sample holder is an optically transparent material including one of an Eppendorf, test tube, glass slide, glass coverslip, or microfluidics chip.

4. The method of claim 1, wherein said wanted objects are sperm and said antibody is a human sperm specific antibody.

5. The method of claim 1, wherein said PCR is one of bulk PCR on multiple sorted objects of a same type, or single cell PCR-based STR analysis.

6. The method of claim 1, further comprising:

releasing the objects from said surface; and

inspecting the wanted objects for quality and purity using one of brightfield microscopy or fluorescent microscopy

7. The method of claim 1, further comprising:

performing said sorting and said PCR-based STR analysis on one of a same or a different chip.

8. The method of claim 1, further comprising:

binding said antigen to a primary antibody; and

binding said primary antibody to a secondary antibody.

9. The method of claim 1, further comprising:

immobilizing ligands on said sample holder;

binding cell surface receptors on a surface of said wanted objects with said ligands.

10. The method of claim 1, further comprising:

binding a ligand to object-specific organomolecules; and

capturing said ligand bound object using an antibody.

11. The method of claim 1, wherein said sample holder contains beads coated with said antibody to bind, one of directly or indirectly, to said wanted objects.

12. The method of claim 11, further comprising:

binding said antigen to a primary antibody; and

binding said primary antibody to a secondary antibody.

wherein said bead is coated with one of Protein A or Protein G, and

wherein said bead recognizes and captures said wanted objects bound to said primary antibody.

13. The method of claim 1, further comprising:

coating one of Protein A or Protein G on a surface of said sample holder;

binding said antigen to a primary antibody; and

binding said primary antibody to a secondary antibody;

wherein said Protein A or Protein G recognizes and captures said wanted objects that are antibody labeled.

14. The method of claim 1, wherein said PCR performing step is compatible with automation and multiplexing.

15. The method of claim 2, wherein said sorting step using antibody-based sorting in conjunction with holographic optical trapping separates maternal and chorionic villi components before PCR-based STR analysis is performed for identification of DNA for paternal testing.

16. The method of claim 1, further comprising:

performing PCR-based STR analysis on the unwanted objects.

17. An apparatus for sorting objects comprising:

a sample holder coated with an antibody, said sample holder containing a sample of wanted objects and unwanted objects, said wanted objects containing an antigen;

wherein said antibody coated on said sample holder binds with said antigen on said wanted objects;

a chip on which said wanted objects are sorted; and

an apparatus for performing PCR-based STR analysis on said wanted objects.

18. The apparatus of claim 17, further comprising:

a holographic optical trapping apparatus which optically traps and sorts said wanted objects from said unwanted objects in said sample.

19. The apparatus of claim 17, wherein said wanted objects are cells and said unwanted objects are contaminants.

20. The apparatus of claim 17, wherein said PCR apparatus performs one of bulk PCR on multiple sorted objects of a same type, or single cell PCR-based STR analysis.

21. The apparatus of claim 17, wherein said chip is a microfluidics chip.

22. The apparatus of claim 17, wherein said sample holder and a substrate on which said apparatus performs said PCR-based STR analysis, are a single chip.

23. The apparatus of claim 18, wherein said apparatus for performing said PCR is automated and multiplexed.

24. The apparatus of claim 18, further comprising:

a plurality of ligands disposed on said sample holder;

wherein cell surface receptors on a surface of said wanted objects bind with said ligands.

25. The apparatus of claim 18, further comprising:

a plurality of ligands disposed on said sample holder;

wherein said ligands are bound to object-specific organomolecules; and

wherein said ligand bound object uses said antibody against said ligand.

26. The apparatus of claim 18, wherein said sample holder is a bead coated with said antibody to bind one of directly or indirectly to said wanted objects.

27. The apparatus of claim 18, wherein said sample holder is coated with one of Protein A or Protein G.

28. The apparatus of claim 26, wherein said antibody further comprises a primary antibody and a secondary antibody;

wherein said primary antibody is bound to said antigen; and

wherein said primary antibody is bound to a secondary antibody;

wherein said bead is coated with one of Protein A or Protein G, and

wherein said bead recognizes and captures said wanted objects bound to said primary antibody.

29. The apparatus of claim 17, wherein PCR-based STR analysis is performed on the unwanted objects.