Methods and compositions for assessing a sample by maldi mass spectrometry

• US 20050178959A1
• Filed: 11/30/2004
• Published: 08/18/2005
• Est. Priority Date: 02/18/2004
• Status: Abandoned Application

• Current Assignee: Agilent Technologies, Inc.
• Original Assignee: Agilent Technologies, Inc.
• Inventors: Hirschberg, David L., Lopez-Avila, Viorica
• Application Number:
  US11/001,493
• Publication Number:
  US 20050178959A1
• Time in Patent Office: Days
• US Class Current: 250/288
• CPC Class Codes: H01J 49/0418 for laser desorption, e.g. matrix-assisted laser desorption/ionisation [MALDI] plates or surface enhanced laser desorption/ionisation [SELDI] plates

Abstract

The invention provides methods for preparing a sample for matrix-assisted laser desorption ionization (MALDI). In general, the methods involve: binding analytes in a sample to capture agents that are bound to matrix that is present in a plurality of wells of a multi-well sample plate, washing any unbound analytes from the matrix, cleaving any bound analyte/capture agent complexes with a MALDI cleavage agent, and depositing the cleavage products on a multi-sample MALDI sample plate. Kits and other compositions are provided for performing the subject methods. The subject invention finds use in methods of simultaneously assessing the presence of several analytes in a single sample, and, as such, the invention finds use in a variety of different medical, research and proteomics applications.

Claims

1. A method for preparing a sample for matrix-assisted laser desorption ionization (MALDI), comprising:

contacting said sample with a matrix attached to capture agents, wherein said matrix is present in a plurality of wells of a multi-well sample plate;

washing said matrix to provide washed matrix;

contacting said washed matrix with a MALDI cleavage agent to produce a solution containing cleavage-agent treated analytes, if present;

separating said solution from said matrix; and

,

depositing said solution onto a MALDI sample plate.

2. The method of claim 1, wherein said solution is deposited in combination with MALDI matrix.

3. The method of claim 1, wherein said separating employs subjecting an exit end of said wells to low atmospheric pressure.

4. The method of claim 1, wherein said matrix contains beads.

5. The method of claim 1, wherein said capture agent is a polypeptide.

6. The method of claim 1, wherein said capture agent is a peptide.

7. The method of claim 1, wherein said capture agent is an antibody.

8. The method of claim 1, wherein said cleavage agent is a chemical or enzymatic cleavage agent

9. The method of claim 8, wherein said enzymatic cleavage agent is an enzyme.

10. The method of claim 8, further comprising crystallizing said solution with MALDI matrix on said MALDI sample plate.

11. The method of claim 1, wherein said capture agent is non-covalently or covalently linked to said porous matrix.

12. The method of claim 1, further comprising ionizing said sample by MALDI.

19. A sample analysis method, comprising:

preparing a set of samples according to the method of claim 1;

ionizing said samples using MALDI;

evaluating the mass of said fragments using mass spectrometry.

20. The method of claim 19, wherein results obtained from said method are compared to results obtained from a control.

21. The method of claim 20, wherein said control method includes a control sample that does not contain any analyte that binds to said capture agent.

22. The method of claim 19, wherein said evaluating includes employing time-of-flight mass spectrometry.

13. A multi-well sample plate having a plurality of wells, at least one well of which comprises:

a matrix attached to analyte/capture agent complexes; and

a MALDI cleavage agent.

14. The multi-well sample plate of claim 13, wherein said MALDI cleavage agent is a chemical cleavage agent

15. The multi-well sample plate of claim 13, wherein said MALDI cleavage agent is an enzymatic cleavage agent

16. The multi-well sample plate of claim 13, wherein said capture agent is a peptide.

17. The multi-well sample plate of claim 13, wherein said capture agent is non-covalently attached to said porous matrix

18. A system for evaluating a sample, comprising:

a multi-well sample plate comprising wells having a matrix attached to a capture agent;

a MALDI cleavage agent; and

a MALDI matrix.