Ligand array assays that include an organic fluid wash step and compositions for practicing the same

• US 20040241666A1
• Filed: 05/30/2003
• Published: 12/02/2004
• Est. Priority Date: 05/30/2003
• Status: Abandoned Application

• Current Assignee: Agilent Technologies, Inc.
• Original Assignee: Agilent Technologies, Inc.
• Inventors: Leproust, Eric M., Amorese, Douglas A., Ke, Winny W.
• Application Number:
  US10/452,767
• Publication Number:
  US 20040241666A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C12Q 1/6837 using probe arrays or probe chips C12Q1/6874 takes precedence

Abstract

Ligand array assays and compositions for use in practicing the same are provided. A feature of the subject methods is that they include a wash step in which the ligand displaying surface of a sample exposed ligand array is washed with organic wash fluid, e.g., propylene carbonate. Also provided are kits for use in practicing the subject methods. The subject methods and kits find use in a variety of ligand array based applications, including genomic and proteomic applications.

Claims

1. A method of determining whether an analyte is present in a sample, said method comprising:

(a) contacting said sample with a surface of a substrate having immobilized thereon a ligand that specifically binds to said analyte;

(b) washing said surface with a an organic wash fluid in which said analyte and ligand therefore are not soluble; and

(c) detecting any resultant binding complexes on said surface to determine whether said analyte is present in said sample.

2. The method according to claim 1, wherein said fluid is a high surface tension fluid.

3. The method according to claim 1, wherein said wash fluid has a surface tension that is at least about 40 mN/m.

4. The method according to claim 1, wherein said wash fluid has a vapor pressure of less than about 10⁻

1 KPa.

5. The method according to claim 4, wherein said wash fluid comprises propylene carbonate.

6. The method according to claim 1, wherein said analyte is a nucleic acid.

7. The method according to claim 1, wherein said ligand is a nucleic acid.

8. The method according to claim 7, wherein said substrate displaying said nucleic acid ligand is a nucleic acid array.

9. The method according to claim 8, wherein said nucleic acid array is an in situ prepared nucleic acid array.

10. The method according to claim 1, wherein said method further comprises at least one additional wash step prior to said wash step (b).

11. The method according to claim 10, wherein said at least one additional wash step comprises washing said substrate surface with an aqueous fluid.

12. The method according to claim 11, wherein said organic wash fluid is miscible with said aqueous fluid.

13. The method according to claim 1, wherein said method is a method of assaying said sample for the presence of two or more distinct analytes.

14. The method according to claim 1, wherein at least a portion of said method is automated.

15. A method comprising transmitting data representing a result obtained by the method according to claim 1, from a first location to a second location.

16. A method according to claim 15, wherein said second location is a remote location.

17. A method comprising receiving data representing a result of a method of claim 1.

18. A kit for performing an assay, said kit comprising:

(a) a high surface tension organic fluid in which nucleic acids are not soluble; and

(b) instructions for using said wash fluid in a method according to claim 1.

19. The kit according to claim 18, wherein said wash fluid has a surface tension that is at least about 40 mN/m.

20. The kit according to claim 18, wherein said wash fluid has a vapor pressure of less than about 10⁻

1 KPa.

21. The kit according to claim 20, wherein said wash fluid comprises propylene carbonate.

22. The kit according to claim 18, wherein said wash fluid comprises at least about 2% (v/v) of a cosolvent.

23. The kit according to claim 18, wherein said kit further comprises an array comprising at least two different ligands.

24. The kit according to claim 23, wherein said ligands are nucleic acid ligands.

25. A substrate produced according to the method of claim 1.

26. The substrate according to claim 25, wherein said substrate is a substrate of a ligand array having two or more distinct ligands immobilized on a surface of said substrate.

27. The substrate according to claim 26, wherein said ligand array is a nucleic acid array.

28. The substrate according to claim 27, wherein said nucleic acid array is an in situ prepared nucleic acid array.